Reduced viscosity protein pharmaceutical formulations

ABSTRACT

Described herein are therapeutic protein formulations comprising N-methyl pyrrolidone (NMP), methods related to reducing viscosity of pharmaceutical formulations and methods related to stabilizing pharmaceutical formulations using NMP.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent ApplicationNo. 62/404,861, filed Oct. 6, 2016, the contents of which areincorporated by reference.

SEQUENCE LISTING

The present application is being filed with a sequence listing inelectronic format. The sequence listing provided as a file titled,“A-2092-WO-PCT_sequence_listing_ST25.txt”, created Oct. 5, 2017, and is284 KB in size. The information in the electronic format of the sequencelisting is incorporated herein by reference in its entirety.

BACKGROUND

Pharmaceutically active proteins, such as antibodies, are frequentlyformulated in liquid solutions, particularly for parenteral injection.For products that need to be administered via a subcutaneous route, forexample use in self-administration, formulations in delivery volumesgreater than 1-2 milliliters are not well tolerated. In such cases,highly concentrated protein formulations are desirable to meet thelimited dose volume. The high dose and small volume requirements of suchadministration means that the protein therapeutic can reachconcentrations of upwards of 100 mg/mL or more. Highly concentratedprotein formulations can pose many challenges to the manufacturabilityand administration of protein therapeutics. One challenge posed by somehighly concentrated protein formulations is increased viscosity. Highviscosity formulations are difficult to handle during manufacturing,including at the bulk and filling stages. High viscosity formulationsare also difficult to draw into a syringe and inject, makingadministration to the patient difficult and unpleasant. There is a needin the pharmaceutical industry to identify compounds that are useful forreducing viscosity of highly concentrated protein formulations, todevelop methods of reducing the viscosity of such formulations, and toprovide pharmaceutical formulations with reduced viscosity.

SUMMARY

Provided is N-methyl pyrrolidone (NMP) at selected concentrations foruse in reducing the viscosity of protein pharmaceutical formulations.NMP can be used with other excipients, such as for example, arginine,lysine, N-acetyl arginine (NAR), and mixtures thereof. Methods forreducing the viscosity of protein pharmaceutical formulations bycombining a high concentration of a therapeutic protein with aviscosity-reducing concentration of NMP are provided herein. Alsoprovided are lyophilized powders comprising a therapeutic protein andNMP, wherein NMP is present at a weight:weight concentration effectiveto reduce viscosity upon reconstitution with a diluent.

Provided herein are methods for reducing the viscosity of a liquidpharmaceutical formulation comprising a therapeutic protein at aconcentration of at least about 100 mg/mL, such as about 100 mg/mL toabout 300 mg/mL (e.g., about 140 mg/mL), such as about 180 mg/mL toabout 230 mg/mL, such as about 210 mg/mL, comprising the step ofcombining the therapeutic protein with a viscosity-reducingconcentration of NMP. In one embodiment the viscosity of the formulationis reduced by at least about 5%. In another embodiment the viscosity ofthe formulation is reduced by at least about 10%. In another embodimentthe viscosity of the formulation is reduced by at least about 20%. Inanother embodiment the viscosity of the formulation is reduced by atleast about 30%. In another embodiment the viscosity of the formulationis reduced by at least about 40%. In another embodiment the viscosity ofthe formulation is reduced by at least about 50%. In another embodimentthe viscosity of the formulation is reduced by at least about 60%. Inanother embodiment the viscosity of the formulation is reduced by atleast about 70%. In another embodiment the viscosity of the formulationis reduced by at least about 80%. In some embodiments, the therapeuticprotein is an antibody (such as an IgG1 or IgG2 monoclonal antibody) orantigen-binding fragment thereof, such as, for example, an anti-PCSK9antibody or antigen-binding fragment thereof. An example of ananti-PCSK9 antibody is evolocumab. In a related embodiment,pharmaceutical formulations produced by such methods are provided. Otherexcipients can be used in combination with NMP, such as arginine,lysine, NAR, and mixtures thereof.

Also provided is a pharmaceutical formulation comprising a therapeuticprotein at a concentration of at least about 100 mg/mL, such as about100 mg/mL to about 300 mg/mL (e.g., about 140 mg/mL), such as about 180mg/mL to about 230 mg/mL, such as about 210 mg/mL, and NMP. In oneembodiment, the concentration of NMP is from about 1.0 mM to about 1000mM. In a related embodiment the concentration of NMP is from about 100mM to about 500 mM. In another related embodiment the concentration ofNMP is from about 300 mM to about 425 mM. In another related embodimentthe concentration of NMP is about 340 mM. In another related embodimentthe concentration of NMP is about 425 mM. In a related embodiment theconcentration of NMP is greater than about 300 mM, greater than about500 mM, greater than about 800 mM, about 1000 mM. Also provided are suchpharmaceutical formulations having a pH between about 4.0 to about 6.0.In a related embodiment, the pH is about 4.8 to about 5.4. In someembodiments, the pH is about 5.0. In yet other related embodiments, thetherapeutic protein is an antibody (such as an IgG1 or IgG2 monoclonalantibody) or antigen-binding fragment thereof, such as, for example, ananti-PCSK9 antibody or antigen-binding fragment thereof. An example ofan anti-PCSK9 antibody is evolocumab.

In one preferred aspect, a pharmaceutical formulation comprises at leastabout 100 mg/mL, such as about 100 mg/mL to about 300 mg/mL (e.g., about140 mg/mL), such as about 180 mg/mL to about 230 mg/mL, such as about210 mg/mL of a therapeutic protein, e.g., an antibody (such as an IgG1or IgG2 monoclonal antibody; an example of an antibody is an anti-PCSK9antibody, such as evolocumab) or an antigen-binding fragment thereof,about 370-600 mM NMP, and about 0.05-0.2% PLURONIC® F-68(polyoxyethylene-polyoxypropylene block copolymer); in related aspects,the pharmaceutical formulation further comprises a buffer, such as about5-20 mM acetate; in yet further aspects, the pharmaceutical formulationhas a pH of about 4.5-5.5. In one embodiment, the pharmaceuticalformulation comprises at least about 100 mg/mL, such as about 100 mg/mLto about 300 mg/mL (e.g., about 140 mg/mL), such as about 180 mg/mL toabout 230 mg/mL, such as about 210 mg/mL of an antibody (such as an IgG1or IgG2 monoclonal antibody) or antigen-binding fragment thereof (e.g.,an anti-PCSK9 antibody, such as evolocumab), about 425 mM NMP, and about0.10% PLURONIC® F-68; in related aspects, the pharmaceutical formulationfurther comprises a buffer, such as about 10 mM acetate; in yet furtheraspects, the pharmaceutical formulation has a pH of about 5.0. Inanother embodiment, the pharmaceutical formulation comprises at leastabout 100 mg/mL of an antibody or (e.g., an anti-PCSK9 antibody, such asevolocumab) or an antigen-binding fragment thereof, about 340 mM NMP,and about 0.10% PLURONIC® F-68; in related aspects, the pharmaceuticalformulation further comprises a buffer, such as about 10 mM acetate; inyet further aspects, the pharmaceutical formulation has a pH of about5.0.

In another preferred aspect, a pharmaceutical formulation comprises atleast about 100 mg/mL, such as about 100 mg/mL to about 300 mg/mL (e.g.,about 140 mg/mL), such as about 180 mg/mL to about 230 mg/mL, such asabout 210 mg/mL of a therapeutic protein, e.g., an antibody (such as anIgG1 or IgG2 monoclonal antibody; an example of an antibody is ananti-PCSK9 antibody, such as evolocumab) or antigen-binding fragmentthereof, about 50-200 mM NMP, about 90-160 mM lysine, and about0.005-0.02% polysorbate 80 (polyoxyethylenesorbitan monooleate, alsoknown as TWEEN® 80); in related aspects, the pharmaceutical formulationfurther comprises a buffer, such as about 5-20 mM acetate; in yetfurther aspects, the pharmaceutical formulation has a pH of about4.5-5.5. In one embodiment, the pharmaceutical formulation comprises atleast about 100 mg/mL of an antibody or (e.g., an anti-PCSK9 antibody,such as evolocumab) or an antigen binding fragment thereof, about 100 mMNMP, about 120 mM lysine, and about 0.01% polysorbate 80; in relatedaspects, the pharmaceutical formulation further comprises a buffer, suchas about 20 mM acetate; in yet further aspects, the pharmaceuticalformulation has a pH of about 5.0.

In another preferred aspect, a pharmaceutical formulation comprises atleast about 100 mg/mL, such as about 100 mg/mL to about 300 mg/mL (e.g.,about 140 mg/mL), such as about 180 mg/mL to about 230 mg/mL, such asabout 210 mg/mL of a therapeutic protein, e.g., an antibody (such as anIgG1 or IgG2 monoclonal antibody; an example of an antibody is ananti-PCSK9 antibody, such as evolocumab) or an antigen-binding fragmentthereof, about 50-200 mM NMP, about 90-160 mM arginine, and about0.005-0.02% polysorbate 80; in related aspects, the pharmaceuticalformulation further comprises a buffer, such as about 10-30 mM acetate;in yet further aspects, the pharmaceutical formulation has a pH of about4.5-5.5.

In one embodiment, the pharmaceutical formulation comprises at leastabout 100 mg/mL, such as about 100 mg/mL to about 300 mg/mL (e.g., about140 mg/mL), such as about 180 mg/mL to about 230 mg/mL, such as about210 mg/mL of an antibody (such as an IgG1 or IgG2 monoclonal antibody;an example of an antibody is an anti-PCSK9 antibody, such as evolocumab)or antigen binding fragment thereof, about 70 mM NMP, about 130 mMarginine, and about 0.01% polysorbate 80; in related embodiments, theformulation further comprises a buffer, such as about 20 mM acetate; inyet further embodiments, the pharmaceutical formulation has a pH ofabout 5.0.

In another preferred aspect, a pharmaceutical formulation comprises atleast about 100 mg/mL of a therapeutic protein, e.g., and antibody(e.g., an anti-PCSK9 antibody, such as evolocumab) or an antigen-bindingfragment thereof, about 50-150 mM NMP, about 90-200 mM N-acetyl arginine(NAR), and about 0.005-0.02% polysorbate 80, and about 5-20 mM acetatehas a pH of 4.5-5.5. In one embodiment, the pharmaceutical formulationcomprises at least about 100 mg/mL of an antibody (e.g., an anti-PCSK9antibody, such as evolocumab), 10 mM acetate, 100 mM NMP, 155 mM NAR,0.01% polysorbate 80, and has a pH of 5.0.

Also provided is a method of preparing a lyophilized powder comprisingthe step of lyophilizing a pharmaceutical formulation as describedabove.

Provided herein is a lyophilized powder comprising a therapeuticprotein, wherein NMP is present at a weight:weight concentrationeffective to reduce viscosity upon reconstitution with a diluent. In oneembodiment NMP is present at a concentration of between about 100 μg/mgtherapeutic protein to about 1 mg/mg therapeutic protein, such as anantibody (such as an IgG1 or IgG2 monoclonal antibody; an example of anantibody is an anti-PCSK9 antibody, such as evolocumab) or anantigen-binding fragment thereof. In a related embodiment, NMP ispresent at a concentration between about 200 μg to about 500 μg/mgtherapeutic protein. Also provided is a method for reconstituting alyophilized powder as described above comprising the step of adding asterile aqueous diluent. Other excipients, such as for example,arginine, lysine, N-acetyl arginine (NAR), and mixtures thereof, can beincluded with NMP.

Also provided are therapeutic proteins that are antibodies (such as IgG1or IgG2 monoclonal antibodies) or antigen-binding fragments thereof.Also provided are formulations or compositions as described abovewherein the therapeutic protein is an antibody or an antigen-bindingfragment thereof. In addition, also provided herein is a lyophilizedpowder as described above wherein the therapeutic protein is an antibodyor an antigen-binding fragment thereof.

For example, therapeutic proteins and antibodies (and antigen-bindingfragments thereof) include Activase® (Alteplase); alirocumab (anti-PCSK9monoclonal antibody designated as H1H316P, see U.S. Pat. No. 8,062,640);Aranesp® (Darbepoetin-alfa), Epogen® (Epoetin alfa, or erythropoietin);Avonex® (Interferon β-Ia); Bexxar® (Tositumomab); Bseron®(Interferon-β); bococizumab (anti-PCSK9 monoclonal antibody designatedas L1L3, see U.S. Pat. No. 8,080,243); Campath® (Alemtuzumab); Dynepo®(Epoetin delta); Velcade® (bortezomib); MLN0002 (anti-α4β7 mAb); MLN1202(anti-CCR2 chemokine receptor mAb); Enbrel® (etanercept); Eprex®(Epoetin alfa); Erbitux® (Cetuximab); evolocumab (anti-PCSK9 monoclonalantibody designated as 21B12, see U.S. Pat. No. 8,030,467); Genotropin®(Somatropin); Herceptin® (Trastuzumab); Humatrope® (somatropin [rDNAorigin] for injection); Humira® (Adalimumab); Infergen® (InterferonAlfacon-1); Natrecor® (nesiritide); Kineret® (Anakinra), Leukine®(Sargamostim); LymphoCide® (Epratuzumab); Benlysta™ (Belimumab);Metalyse® (Tenecteplase); Mircera® (methoxy polyethylene glycol-epoetinβ); Mylotarg® (Gemtuzumab ozogamicin); Raptiva® (efalizumab); Cimzia®(certolizumab pegol); Soliris™ (Eculizumab); Pexelizumab (Anti-C5Complement); MEDI-524 (Numax®); Lucentis® (Ranibizumab); Edrecolomab(,Panorex®); Trabio® (lerdelimumab); TheraCim hR3 (Nimotuzumab);Omnitarg (Pertuzumab, 2C4); Osidem® (IDM-I); OvaRex® (B43.13); Nuvion®(visilizumab); Cantuzumab mertansine (huC242-DMI); NeoRecormon® (Epoetinβ); Neumega® (Oprelvekin); Neulasta® (Pegylated filgastrim, pegylatedG-CSF, pegylated hu-Met-G-CSF); Neupogen® (Filgrastim); Orthoclone OKT3®(Muromonab-CD3), Procrit® (Epoetin alfa); Remicade® (Infliximab),Reopro® (Abciximab), Actemra® (anti-IL6 Receptor mAb), Avastin®(Bevacizumab), HuMax-CD4 (zanolimumab), Rituxan® (Rituximab); Tarceva®(Erlotinib); Roferon-A®-(Interferon alfa-2a); Simulect® (Basiliximab);Stelara™ (Ustekinumab); Prexige® (lumiracoxib); Synagis® (Palivizumab);146B7-CHO (anti-IL15 antibody, see U.S. Pat. No. 7,153,507), Tysabri®(Natalizumab); Valortim® (MDX-1303, anti-B. anthracis Protective AntigenmAb); ABthrax™; Vectibix® (Panitumumab); Xolair® (Omalizumab), ETI211(anti-MRSA mAb), IL-I Trap (the Fc portion of human IgGI and theextracellular domains of both IL-I receptor components (the Type Ireceptor and receptor accessory protein)), VEGF Trap (Ig domains ofVEGFRI fused to IgGI Fc), Zenapax® (Daclizumab); Zenapax® (Daclizumab),Zevalin® (Ibritumomab tiuxetan), Zetia (ezetimibe), Atacicept (TACI-Ig),anti-α4β7 mAb (vedolizumab); galiximab (anti-CD80 monoclonal antibody),anti-CD23 mAb (lumiliximab); BR2-Fc (huBR3/huFc fusion protein, solubleBAFF antagonist); Simponi™ (Golimumab); Mapatumumab (human anti-TRAILReceptor-1 mAb); Ocrelizumab (anti-CD20 human mAb); HuMax-EGFR(zalutumumab); M200 (Volociximab, anti-α5β1 integrin mAb); MDX-010(Ipilimumab, anti-CTLA-4 mAb and VEGFR-I (IMC-18F1); anti-BR3 mAb;anti-Clostridium difficile Toxin A and Toxin B C mAbs MDX-066 (CDA-I)and MDX-1388); anti-CD22 dsFv-PE38 conjugates (CAT-3888 and CAT-8015);anti-CD25 mAb (HuMax-TAC); Adecatumumab (MT201, anti-EpCAM-CD326 mAb);MDX-060, SGN-30, SGN-35 (anti-CD30 mAbs); MDX-1333 (anti-IFNAR); HuMaxCD38 (anti-CD38 mAb); anti-CD40L mAb; anti-Cripto mAb; anti-CTGFIdiopathic Pulmonary Fibrosis Phase I Fibrogen (FG-3019); anti-CTLA4mAb; anti-eotaxinI mAb (CAT-213); anti-FGF8 mAb; anti-ganglioside GD2mAb; anti-ganglioside GM2 mAb; anti-GDF-8 human mAb (MYO-029);anti-GM-CSF Receptor mAb (CAM-3001); anti-HepC mAb (HuMax HepC);MEDI-545, MDX-1103 (anti-IFNα mAb); anti-IGFIR mAb; anti-IGF-IR mAb(HuMax-Inflam); anti-IL12/IL23p40 mAb (Briakinumab); anti-IL-23p19 mAb(LY2525623); anti-IL13 mAb (CAT-354); anti-IL-17 mAb (AIN457);anti-IL2Ra mAb (HuMax-TAC); anti-IL5 Receptor mAb; anti-integrinreceptors mAb (MDX-018, CNTO 95); anti-IPIO Ulcerative Colitis mAb(MDX-1100); anti-LLY antibody; BMS-66513; anti-Mannose Receptor/hCGβ mAb(MDX-1307); anti-mesothelin dsFv-PE38 conjugate (CAT-5001); anti-PDImAb(MDX-1 106 (ONO-4538)); anti-PDGFRα antibody (IMC-3G3); anti-TGFβ mAb(GC-1008); anti-TRAIL Receptor-2 human mAb (HGS-ETR2); anti-TWEAK mAb;anti-VEGFR/Flt-1 mAb; anti-ZP3 mAb (HuMax-ZP3); NVS Antibody #1; and NVSAntibody #2.

Additional examples of antibodies (and antigen-binding fragmentsthereof) include the antibodies shown in Table A. Other examples ofantibodies (and antigen-binding fragments thereof) include abagovomab,abciximab, actoxumab, adalimumab, afelimomab, afutuzumab, alacizumab,alacizumab pegol, ald518, alemtuzumab, alemtuzumab, alirocumab,altinumab, altumomab, amatuximab, anatumomab mafenatox, anrukinzumab,apolizumab, arcitumomab, aselizumab, atlizumab, atorolimiumab,bapineuzumab, basiliximab, bavituximab, bectumomab, belimumab,benralizumab, bertilimumab, besilesomab, bevacizumab, bezlotoxumab,biciromab, bivatuzumab, bivatuzumab mertansine, blinatumomab,blosozumab, brentuximab vedotin, briakinumab, brodalumab, canakinumab,cantuzumab mertansine, cantuzumab mertansine, caplacizumab, capromabpendetide, carlumab, catumaxomab, cc49, cedelizumab, certolizumab pegol,cetuximab, citatuzumab bogatox, cixutumumab, clazakizumab, clenoliximab,clivatuzumab tetraxetan, conatumumab, cr6261, crenezumab, dacetuzumab,daclizumab, dalotuzumab, daratumumab, demcizumab, denosumab, detumomab,dorlimomab aritox, drozitumab, duligotumab, dupilumab, ecromeximab,eculizumab, edobacomab, edrecolomab, efalizumab, efungumab, elotuzumab,elsilimomab, enavatuzumab, enlimomab pegol, enokizumab, enokizumab,enoticumab, enoticumab, ensituximab, epitumomab cituxetan, epratuzumab,erenumab, erlizumab, ertumaxomab, etaracizumab, etrolizumab, evolocumab,exbivirumab, exbivirumab, fanolesomab, faralimomab, farletuzumab,fasinumab, fbta05, felvizumab, fezakinumab, ficlatuzumab, figitumumab,flanvotumab, fontolizumab, foralumab, foravirumab, fresolimumab,fulranumab, futuximab, galiximab, ganitumab, gantenerumab, gavilimomab,gemtuzumab ozogamicin, gevokizumab, girentuximab, glembatumumab vedotin,golimumab, gomiliximab, gs6624, ibalizumab, ibritumomab tiuxetan,icrucumab, igovomab, imciromab, i mgatuzumab, inclacumab, indatuximabravtansine, infliximab, inolimomab, inotuzumab ozogamicin, intetumumab,ipilimumab, iratumumab, itolizumab, ixekizumab, keliximab, labetuzumab,lebrikizumab, lemalesomab, lerdelimumab, lexatumumab, libivirumab,ligelizumab, lintuzumab, lirilumab, lorvotuzumab mertansine,lucatumumab, lumiliximab, mapatumumab, maslimomab, matuzumab,mavrilimumab, mepolizumab, metelimumab, milatuzumab, minretumomab,mitumomab, mogamulizumab, morolimumab, motavizumab, moxetumomabpasudotox, muromonab-cd3, nacolomab tafenatox, namilumab, naptumomabestafenatox, narnatumab, natalizumab, nebacumab, necitumumab,nerelimomab, nesvacumab, nimotuzumab, nivolumab, nofetumomab merpentan,ocaratuzumab, ocrelizumab, odulimomab, ofatumumab, olaratumab,olokizumab, omalizumab, onartuzumab, oportuzumab monatox, oregovomab,orticumab, otelixizumab, oxelumab, ozanezumab, ozoralizumab,pagibaximab, palivizumab, palivizumab, panitumumab, panobacumab,parsatuzumab, pascolizumab, pateclizumab, patritumab, pemtumomab,perakizumab, pertuzumab, pexelizumab, pidilizumab, pintumomab,placulumab, ponezumab, priliximab, pritumumab, PRO 140, quilizumab,racotumomab, radretumab, rafivirumab, ramucirumab, ranibizumab,raxibacumab, regavirumab, reslizumab, rilotumumab, rituximab,robatumumab, roledumab, romosozumab, rontalizumab, rovelizumab,ruplizumab, samalizumab, sarilumab, satumomab pendetide, secukinumab,sevirumab, sibrotuzumab, sifalimumab, siltuximab, simtuzumab,siplizumab, sirukumab, solanezumab, solitomab, sonepcizumab, sontuzumab,stamulumab, sulesomab, suvizumab, tabalumab, tacatuzumab tetraxetan,tadocizumab, talizumab, tanezumab, taplitumomab paptox, tefibazumab,tefibazumab, telimomab aritox, telimomab aritox, tenatumomab,tenatumomab, teneliximab, teplizumab, teprotumumab, TGN1412,ticilimumab, tigatuzumab, tildrakizumab, TNX-650, tocilizumab,tocilizumab, toralizumab, tositumomab, tralokinumab, trastuzumab,TRBS07, tregalizumab, tremelimumab, tremelimumab, tucotuzumabcelmoleukin, tuvirumab, ublituximab, urelumab, urtoxazumab, ustekinumab,vapaliximab, vatelizumab, vedolizumab, veltuzumab, vepalimomab,vesencumab, visilizumab, volociximab, vorsetuzumab mafodotin, votumumab,zalutumumab, zanolimumab, zatuximab, ziralimumab, and zolimomab aritox.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1B. Changes in solution viscosity at 25° C. FIG. 1A: Measuredsolution viscosity as a function of protein concentration in presence ofexcipients (●) 1000 mM NMP, (Δ) 340 mM NMP, (◯) 1000 mM DMA, (▾) 1000 mMDMSO, (□) 170 mM arginine and (▪) 220 mM proline over a range ofconcentrations 110 mg/mL to 220 mg/mL. Each data point represents anaverage of two independent readings. Error bars are displayed as 2 timesstandard deviation of the measurements. FIG. 1B: Measured solutionviscosity as a function of increasing NMP concentrations at 210 mg/mLevolocumab

FIG. 2. Changes in solution viscosity at 25° C. Measured solutionviscosity as a function of protein concentration in presence ofexcipients (●) 130 mM Arginine, 70 mM NMP, (▪) 340 mM NMP, (◯) 170 mMArginine and (▾) 220 mM Proline over a range of concentrations 110 mg/mLto 220 mg/mL. Each data point represents an average of two independentreadings. Error bars are displayed as 2 times standard deviation of themeasurements.

FIG. 3. Evolocumab viscosity as a function of the concentration ofdifferent excipients.

FIG. 4. Effect of 4% NMP on a variety of mAbs. There were three IgG2antibodies tested (IgG2 Ab A, IgG2 Ab B, and IgG2 Ab C), and three IgG1antibodies tested (IgG1 Ab A, IgG1 Ab B, and IgG1 Ab C). It was seenthat addition of 4% NMP (striped bar for each mAb) had significantlylower viscosity compared to no excipient for 5 out of the 6 molecules.The molecule where NMP did not have a significant effect (same ascontrol) was IgG1 Ab B. It is possible that this was due to the presenceof Fab glycosylation for IgG1 Ab B which was absent from all the othermolecules tested.

DETAILED DESCRIPTION

Reducing the viscosity of high concentration therapeutic proteinformulations is of interest to the pharmaceutical industry. NMP wasdiscovered to reduce the viscosity of liquid formulations. Suchformulations comprising NMP were also found to have lowered viscosity inthe presence of optional, additional excipients, such as for example,arginine, lysine, N-acetyl arginine (NAR), and mixtures thereof.Provided herein is NMP at selected concentrations for use in reducingthe viscosity of protein formulations. Methods for reducing theviscosity of protein formulations by combining the therapeutic proteinwith a viscosity-reducing concentration of NMP is provided herein. Alsoprovided is lyophilized powder comprising a therapeutic protein and NMP,wherein NMP is present at a weight:weight concentration effective toreduce viscosity upon reconstitution with a diluent.

Unless otherwise required by context, singular terms shall includepluralities and plural terms shall include the singular. Generally,nomenclatures used in connection with, and techniques of, cell andtissue culture, molecular biology, immunology, microbiology, geneticsand protein and nucleic acid chemistry and hybridization describedherein are those well-known and commonly used in the art. The disclosedmethods and techniques are generally performed according to conventionalmethods well-known in the art and as described in various general andmore specific references that are cited and discussed throughout thepresent specification unless otherwise indicated. See, e.g., Sambrook etal., Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring HarborLaboratory Press, Cold Spring Harbor, N.Y. (2001) and Ausubel et al.,Current Protocols in Molecular Biology, Greene Publishing Associates(1992), and Harlow and Lane, Antibodies: A Laboratory Manual, ColdSpring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1990).Enzymatic reactions and purification techniques are performed accordingto manufacturer's specifications, as commonly accomplished in the art oras described herein. The terminology used in connection with, and thelaboratory procedures and techniques of, analytical chemistry, syntheticorganic chemistry, and medicinal and pharmaceutical chemistry describedherein are those well-known and commonly used in the art. Standardtechniques can be used for chemical syntheses, chemical analyses,pharmaceutical preparation, formulation, and delivery, and treatment ofpatients. Stated ranges include all values in between the endpoints ofthe range.

All patents and other publications identified are expressly incorporatedherein by reference in their entirety for the purpose of describing anddisclosing, for example, the methodologies described in suchpublications that might be used in connection with the described.

NMP (N-methyl pyrrolidone) is an organic pharmaceutical excipient thathas been employed in both parenteral as well as oral formulations. See,e.g., “Review of Pharmaceutical Applications of N-Methyl-2-Pyrrolidone.”Jouyban, A; Fakhree M A A and Shayanfar A J. Pharm. Pharmaceut. Sci.13(4) 524-535.

The terms “polypeptide” and “protein” are used interchangeably herein.Exemplary polypeptides contemplated for use in the stable pharmaceuticalformulations disclosed herein include antibodies, peptibodies,immunoglobulin-like proteins, non-antibody proteins andnon-immunoglobulin-like proteins. Analogs of naturally occurringproteins are contemplated for inclusion in the disclosed formulations,including polypeptides with modified glycosylation, polypeptides withoutglycosylation (unglycosylated). As used herein, “analogs” refers to anamino acid sequence that has insertions, deletions or substitutionsrelative to the parent sequence, while still substantially maintainingthe biological activity of the parent sequence, as determined usingbiological assays known to one of skill in the art. The disclosedformulations can also include derivatives of naturally occurring oranalog polypeptides which have been chemically modified, for example, toattach water soluble polymers (e.g., PEGylated), radionuclides, or otherdiagnostic or targeting or therapeutic moieties.

Antibodies can be formulated using the methods and compositionsdisclosed herein. As used herein, the term “antibody” includes fullyassembled antibodies, monoclonal antibodies (including human, humanizedor chimeric antibodies), polyclonal antibodies, multispecific antibodies(e.g., bispecific antibodies), maxibody, and antibody fragments that canbind antigen (e.g., Fab′, F′(ab)2, Fv, single chain antibodies,diabodies), comprising complementarity determining regions (CDRs) of theforegoing as long as they exhibit the desired biological activity.

Peptibodies, molecules comprising an antibody Fc domain attached to atleast one antigen-binding peptide, are generally described in PCTpublication WO 00/24782. Immunoglobulin-like proteins, members of theimmunoglobulin superfamily, contain one or more immunoglobulin-likedomains which fold in structures similar to portions of the antibodyvariable region.

Proteins, including those that bind to one or more of the following, areuseful in the disclosed compositions and methods. These include CDproteins including, but not limited to, CD3, CD4, CD8, CD19, CD20, CD22,CD30, and CD34; including those that interfere with receptor binding.HER receptor family proteins, including HER2, HER3, HER4, and the e.g.,F receptor. Cell adhesion molecules, for example, LFA-I, Mol, pI50, 95,VLA-4, ICAM-I, VCAM, and α v/β 3 integrin. Growth factors, including butnot limited to, vascular endothelial growth factor (“VEGF”), growthhormone, thyroid stimulating hormone, follicle stimulating hormone,luteinizing hormone, growth hormone releasing factor, parathyroidhormone, Mullerian-inhibiting substance, human macrophage inflammatoryprotein (MIP-I-α), erythropoietin (EPO), nerve growth factor, such asNGF-β, platelet-derived growth factor (PDGF), fibroblast growth factors,including, for instance, aFGF and bFGF, epidermal growth factor (EGF),transforming growth factors (TGF), including, among others, TGF-α andTGF-β, including TGF-βI, TGF-β2, TGF-β3, TGF-β4, or TGF-β5, insulin-likegrowth factors-I and -II (IGF-I and IGF-II), des(I-3)-IGF-I (brainIGF-I), and osteoinductive factors. Insulins and insulin-relatedproteins, including but not limited to, insulin, insulin A-chain,insulin B-chain, proinsulin, and insulin-like growth factor bindingproteins. Coagulation and coagulation-related proteins, such as, amongothers, factor VIII, tissue factor, von Willebrands factor, protein C,α-1-antitrypsin, plasminogen activators, such as urokinase and tissueplasminogen activator (“t-PA”), bombazine, thrombin, and thrombopoietin;(vii) other blood and serum proteins, including but not limited to,albumin, IgE, and blood group antigens. Colony stimulating factors andreceptors thereof, including the following, among others, M-CSF, GM-CSF,and G-CSF, and receptors thereof, such as CSF-1 receptor (c-fms).Receptors and receptor-associated proteins, including, for example,flk2/flt3 receptor, obesity (OB) receptor, LDL receptor, growth hormonereceptors, thrombopoietin receptors (“TPO-R,” “c-mpl”), glucagonreceptors, interleukin receptors, interferon receptors, T-cellreceptors, stem cell factor receptors, such as c-Kit, and otherreceptors listed herein. Receptor ligands, including, for example,OX40L, the ligand for the OX40 receptor. Neurotrophic factors, includingbut not limited to, bone-derived neurotrophic factor (BDNF) andneurotrophin-3, -4, -5, or -6 (NT-3, NT-4, NT-5, or NT-6). RelaxinA-chain, relaxin B-chain, and prorelaxin; interferons and interferonreceptors, including for example, interferon-α, -β, and -γ, and theirreceptors. Interleukins and interleukin receptors, including but notlimited to, IL-I to IL-33 and IL-I to IL-33 receptors, such as the IL-8receptor, among others. Viral antigens, including but not limited to, anAIDS envelope viral antigen. Lipoproteins, calcitonin, glucagon, atrialnatriuretic factor, lung surfactant, tumor necrosis factor-α and -β,enkephalinase, RANTES (regulated on activation normally T-cell expressedand secreted), mouse gonadotropin-associated peptide, DNAse, inhibin,and activin. Integrin, protein A or D, rheumatoid factors, immunotoxins,bone morphogenetic protein (BMP), superoxide dismutase, surface membraneproteins, decay accelerating factor (DAF), AIDS envelope, transportproteins, homing receptors, addressins, regulatory proteins,immunoadhesins, antibodies. Myostatins, TALL proteins, including TALL-I,amyloid proteins, including but not limited to, amyloid-β proteins,thymic stromal lymphopoietins (“TSLP”), RANK ligand (“OPGL”), c-kit, TNFreceptors, including TNF Receptor Type 1, TRAIL-R2, angiopoietins, andbiologically active fragments or analogs or variants of any of theforegoing.

Exemplary proteins and antibodies include Activase® (Alteplase);alirocumab (anti-PCSK9 monoclonal antibody designated as H1H316P, seeU.S. Pat. No. 8,062,640); Aranesp® (Darbepoetin-alfa), Epogen® (Epoetinalfa, or erythropoietin); Avonex® (Interferon β-Ia); Bexxar®(Tositumomab); Bseron® (Interferon-β); bococizumab (anti-PCSK9monoclonal antibody designated as L1L3, see U.S. Pat. No. 8,080,243);Campath® (Alemtuzumab); Dynepo® (Epoetin delta); Velcade® (bortezomib);MLN0002 (anti-α4β7 mAb); MLN1202 (anti-CCR2 chemokine receptor mAb);Enbrel® (etanercept); Eprex® (Epoetin alfa); Erbitux® (Cetuximab);evolocumab (anti-PCSK9 monoclonal antibody designated as 21B12, see U.S.Pat. No. 8,030,467; heavy chain: SEQ ID NO:104; light chain: SEQ IDNO:105); Genotropin® (Somatropin); Herceptin® (Trastuzumab); Humatrope®(somatropin [rDNA origin] for injection); Humira® (Adalimumab);Infergen® (Interferon Alfacon-1); Natrecor® (nesiritide); Kineret®(Anakinra), Leukine® (Sargamostim); LymphoCide® (Epratuzumab); Benlysta™(Belimumab); Metalyse® (Tenecteplase); Mircera® (methoxy polyethyleneglycol-epoetin β); Mylotarg® (Gemtuzumab ozogamicin); Raptiva®(efalizumab); Cimzia® (certolizumab pegol); Soliris™ (Eculizumab);Pexelizumab (Anti-C5 Complement); MEDI-524 (Numax®); Lucentis®(Ranibizumab); Edrecolomab (,Panorex®); Trabio® (lerdelimumab); TheraCimhR3 (Nimotuzumab); Omnitarg (Pertuzumab, 2C4); Osidem® (IDM-I); OvaRex®(B43.13); Nuvion® (visilizumab); Cantuzumab mertansine (huC242-DMI);NeoRecormon® (Epoetin β); Neumega® (Oprelvekin); Neulasta® (Pegylatedfilgastrim, pegylated G-CSF, pegylated hu-Met-G-CSF); Neupogen®(Filgrastim); Orthoclone OKT3® (Muromonab-CD3), Procrit® (Epoetin alfa);Remicade® (Infliximab), Reopro® (Abciximab), Actemra® (anti-IL6 ReceptormAb), Avastin® (Bevacizumab), HuMax-CD4 (zanolimumab), Rituxan®(Rituximab); Tarceva® (Erlotinib); Roferon-A®-(Interferon alfa-2a);Simulect® (Basiliximab); Stelara™ (Ustekinumab); Prexige® (lumiracoxib);Synagis® (Palivizumab); 146B7-CHO (anti-IL15 antibody, see U.S. Pat. No.7,153,507), Tysabri® (Natalizumab); Valortim® (MDX-1303, anti-B.anthracis Protective Antigen mAb); ABthrax™; Vectibix® (Panitumumab);Xolair® (Omalizumab), ETI211 (anti-MRSA mAb), IL-I Trap (the Fc portionof human IgGI and the extracellular domains of both IL-I receptorcomponents (the Type I receptor and receptor accessory protein)), VEGFTrap (Ig domains of VEGFRI fused to IgGI Fc), Zenapax® (Daclizumab);Zenapax® (Daclizumab), Zevalin® (Ibritumomab tiuxetan), Zetia(ezetimibe), Atacicept (TACI-Ig), anti-α4β7 mAb (vedolizumab); galiximab(anti-CD80 monoclonal antibody), anti-CD23 mAb (lumiliximab); BR2-Fc(huBR3/huFc fusion protein, soluble BAFF antagonist); Simponi™(Golimumab); Mapatumumab (human anti-TRAIL Receptor-1 mAb); Ocrelizumab(anti-CD20 human mAb); HuMax-EGFR (zalutumumab); M200 (Volociximab,anti-α5β1 integrin mAb); MDX-010 (Ipilimumab, anti-CTLA-4 mAb andVEGFR-I (IMC-18F1); anti-BR3 mAb; anti-Clostridium difficile Toxin A andToxin B C mAbs MDX-066 (CDA-I) and MDX-1388); anti-CD22 dsFv-PE38conjugates (CAT-3888 and CAT-8015); anti-CD25 mAb (HuMax-TAC);Adecatumumab (MT201, anti-EpCAM-CD326 mAb); MDX-060, SGN-30, SGN-35(anti-CD30 mAbs); MDX-1333 (anti-IFNAR); HuMax CD38 (anti-CD38 mAb);anti-CD40L mAb; anti-Cripto mAb; anti-CTGF Idiopathic Pulmonary FibrosisPhase I Fibrogen (FG-3019); anti-CTLA4 mAb; anti-eotaxinI mAb (CAT-213);anti-FGF8 mAb; anti-ganglioside GD2 mAb; anti-ganglioside GM2 mAb;anti-GDF-8 human mAb (MYO-029); anti-GM-CSF Receptor mAb (CAM-3001);anti-HepC mAb (HuMax HepC); MEDI-545, MDX-1103 (anti-IFNα mAb);anti-IGFIR mAb; anti-IGF-IR mAb (HuMax-Inflam); anti-IL12/IL23p40 mAb(Briakinumab); anti-IL-23p19 mAb (LY2525623); anti-IL13 mAb (CAT-354);anti-IL-17 mAb (AIN457); anti-IL2Ra mAb (HuMax-TAC); anti-IL5 ReceptormAb; anti-integrin receptors mAb (MDX-018, CNTO 95); anti-IPIOUlcerative Colitis mAb (MDX-1100); anti-LLY antibody; BMS-66513;anti-Mannose Receptor/hCGβ mAb (MDX-1307); anti-mesothelin dsFv-PE38conjugate (CAT-5001); anti-PDImAb (MDX-1 106 (ONO-4538)); anti-PDGFRαantibody (IMC-3G3); anti-TGFβ mAb (GC-1008); anti-TRAIL Receptor-2 humanmAb (HGS-ETR2); anti-TWEAK mAb; anti-VEGFR/Flt-1 mAb; anti-ZP3 mAb(HuMax-ZP3); NVS Antibody #1; and NVS Antibody #2.

Examples of antibodies suitable for the methods and pharmaceuticalformulations include the antibodies shown in Table A. Other examples ofsuitable antibodies include abagovomab, abciximab, actoxumab,adalimumab, afelimomab, afutuzumab, alacizumab, alacizumab pegol,ald518, alemtuzumab, alemtuzumab, alirocumab, altinumab, altumomab,amatuximab, anatumomab mafenatox, anrukinzumab, apolizumab, arcitumomab,aselizumab, atlizumab, atorolimiumab, bapineuzumab, basiliximab,bavituximab, bectumomab, belimumab, benralizumab, bertilimumab,besilesomab, bevacizumab, bezlotoxumab, biciromab, bivatuzumab,bivatuzumab mertansine, blinatumomab, blosozumab, brentuximab vedotin,briakinumab, brodalumab, canakinumab, cantuzumab mertansine, cantuzumabmertansine, caplacizumab, capromab pendetide, carlumab, catumaxomab,cc49, cedelizumab, certolizumab pegol, cetuximab, citatuzumab bogatox,cixutumumab, clazakizumab, clenoliximab, clivatuzumab tetraxetan,conatumumab, cr6261, crenezumab, dacetuzumab, daclizumab, dalotuzumab,daratumumab, demcizumab, denosumab, detumomab, dorlimomab aritox,drozitumab, duligotumab, dupilumab, ecromeximab, eculizumab, edobacomab,edrecolomab, efalizumab, efungumab, elotuzumab, elsilimomab,enavatuzumab, enlimomab pegol, enokizumab, enokizumab, enoticumab,enoticumab, ensituximab, epitumomab cituxetan, epratuzumab, erenumab,erlizumab, ertumaxomab, etaracizumab, etrolizumab, evolocumab,exbivirumab, exbivirumab, fanolesomab, faralimomab, farletuzumab,fasinumab, fbta05, felvizumab, fezakinumab, ficlatuzumab, figitumumab,flanvotumab, fontolizumab, foralumab, foravirumab, fresolimumab,fulranumab, futuximab, galiximab, ganitumab, gantenerumab, gavilimomab,gemtuzumab ozogamicin, gevokizumab, girentuximab, glembatumumab vedotin,golimumab, gomiliximab, gs6624, ibalizumab, ibritumomab tiuxetan,icrucumab, igovomab, imciromab, i mgatuzumab, inclacumab, indatuximabravtansine, infliximab, inolimomab, inotuzumab ozogamicin, intetumumab,ipilimumab, iratumumab, itolizumab, ixekizumab, keliximab, labetuzumab,lebrikizumab, lemalesomab, lerdelimumab, lexatumumab, libivirumab,ligelizumab, lintuzumab, lirilumab, lorvotuzumab mertansine,lucatumumab, lumiliximab, mapatumumab, maslimomab, matuzumab,mavrilimumab, mepolizumab, metelimumab, milatuzumab, minretumomab,mitumomab, mogamulizumab, morolimumab, motavizumab, moxetumomabpasudotox, muromonab-cd3, nacolomab tafenatox, namilumab, naptumomabestafenatox, narnatumab, natalizumab, nebacumab, necitumumab,nerelimomab, nesvacumab, nimotuzumab, nivolumab, nofetumomab merpentan,ocaratuzumab, ocrelizumab, odulimomab, ofatumumab, olaratumab,olokizumab, omalizumab, onartuzumab, oportuzumab monatox, oregovomab,orticumab, otelixizumab, oxelumab, ozanezumab, ozoralizumab,pagibaximab, palivizumab, palivizumab, panitumumab, panobacumab,parsatuzumab, pascolizumab, pateclizumab, patritumab, pemtumomab,perakizumab, pertuzumab, pexelizumab, pidilizumab, pintumomab,placulumab, ponezumab, priliximab, pritumumab, PRO 140, quilizumab,racotumomab, radretumab, rafivirumab, ramucirumab, ranibizumab,raxibacumab, regavirumab, reslizumab, rilotumumab, rituximab,robatumumab, roledumab, romosozumab, rontalizumab, rovelizumab,ruplizumab, samalizumab, sarilumab, satumomab pendetide, secukinumab,sevirumab, sibrotuzumab, sifalimumab, siltuximab, simtuzumab,siplizumab, sirukumab, solanezumab, solitomab, sonepcizumab, sontuzumab,stamulumab, sulesomab, suvizumab, tabalumab, tacatuzumab tetraxetan,tadocizumab, talizumab, tanezumab, taplitumomab paptox, tefibazumab,tefibazumab, telimomab aritox, telimomab aritox, tenatumomab,tenatumomab, teneliximab, teplizumab, teprotumumab, TGN1412,ticilimumab, tigatuzumab, tildrakizumab, TNX-650, tocilizumab,tocilizumab, toralizumab, tositumomab, tralokinumab, trastuzumab,TRBS07, tregalizumab, tremelimumab, tremelimumab, tucotuzumabcelmoleukin, tuvirumab, ublituximab, urelumab, urtoxazumab, ustekinumab,vapaliximab, vatelizumab, vedolizumab, veltuzumab, vepalimomab,vesencumab, visilizumab, volociximab, vorsetuzumab mafodotin, votumumab,zalutumumab, zanolimumab, zatuximab, ziralimumab, and zolimomab aritox.

TABLE A Exemplary antibodies LC HC Target HC Type LC SEQ SEQ (informalname) (including allotypes) Type pl ID NO ID NO anti-amyloid IgG1 (f)(R; EM) Kappa 9.0 18 19 GMCSF (247) IgG2 Kappa 8.7 20 21 CGRPR IgG2Lambda 8.6 22 23 RANKL IgG2 Kappa 8.6 24 25 Sclerostin (27H6) IgG2 Kappa6.6 26 27 IL-1R1 IgG2 Kappa 7.4 28 29 Myostatin IgG1 (z) (K; EM) Kappa8.7 30 31 B7RP1 IgG2 Kappa 7.7 32 33 Amyloid IgG1 (za) (K; DL) Kappa 8.734 35 GMCSF (3.112) IgG2 Kappa 8.8 36 37 CGRP (32H7) IgG2 Kappa 8.7 3839 CGRP (3B6.2) IgG2 Lambda 8.6 40 41 PCSK9 (8A3.1) IgG2 Kappa 6.7 42 43PCSK9 (492) IgG2 Kappa 6.9 44 45 CGRP IgG2 Lambda 8.8 46 47 HepcidinIgG2 Lambda 7.3 48 49 TNFR p55) IgG2 Kappa 8.2 50 51 OX40L IgG2 Kappa8.7 52 53 HGF IgG2 Kappa 8.1 54 55 GMCSF IgG2 Kappa 8.1 56 57 Glucagon RIgG2 Kappa 8.4 58 59 GMCSF (4.381) IgG2 Kappa 8.4 60 61 Sclerostin(13F3) IgG2 Kappa 7.8 62 63 CD-22 IgG1 (f) (R; EM) Kappa 8.8 64 65 INFgRIgG1 (za) (K; DL) Kappa 8.8 66 67 Ang2 IgG2 Kappa 7.4 68 69 TRAILR2 IgG1(f) (R; EM) Kappa 8.7 70 71 EGFR IgG2 Kappa 6.8 72 73 IL-4R IgG2 Kappa8.6 74 75 IL-15 IgG1 (f) (R; EM) Kappa 8.8 76 77 IGF1R IgG1 (za) (K; DL)Kappa 8.6 78 79 IL-17R IgG2 Kappa 8.6 80 81 Dkk1 (6.37.5) IgG2 Kappa 8.282 83 Sclerostin IgG2 Kappa 7.4 84 85 TSLP IgG2 Lambda 7.2 86 87 Dkk1(11H10) IgG2 Kappa 8.2 88 89 PCSK9 IgG2 Lambda 8.1 90 91 GIPR (2G10.006)IgG1 (z) (K; EM) Kappa 8.1 92 93 Activin IgG2 Lambda 7.0 94 95Sclerostin (2B8) IgG2 Lambda 6.7 96 97 Sclerostin IgG2 Kappa 6.8 98 99c-fms IgG2 Kappa 6.6 100 101 α4β7 IgG2 Kappa 6.5 102 103 HC—antibodyheavy chain; LC—antibody light chain.

“High protein formulation,” “high protein pharmaceutical formulation,”and the like means a formulation that has a protein concentration ofabout 100 mg/mL or greater, such as 100 mg/mL to about 1,000 mg/mL.Exemplary protein concentrations in the formulation can range from about100 mg/mL to about 300 mg/mL, about 120 mg/mL to about 270 mg/mL, fromabout 140 mg/mL to about 255 mg/mL, from about 140 mg/mL to about 240mg/mL, or from about 180 mg/mL to about 230 mg/mL, or alternatively fromabout 190 mg/mL to about 220 mg/mL. The concentration of protein dependson the end use of the pharmaceutical formulation and can be easilydetermined by a person of skill in the art. Particularly contemplatedconcentrations of protein are at least about 140, 145, 150, 155, 160,165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, and 230mg/mL and including all values in between.

As used herein, “pharmaceutical formulation” is a sterile composition ofa pharmaceutically active drug, such as a biologically active protein,that is suitable for parenteral administration (including but notlimited to, intravenous, intramuscular, subcutaneous, aerosolized,intrapulmonary, intranasal or intrathecal) to a patient in need thereofand includes only pharmaceutically acceptable excipients, diluents, andother additives deemed safe by the Federal Drug Administration or otherforeign national authorities. Pharmaceutical formulations includeliquid, e.g., aqueous, solutions that can be directly administered, andlyophilized powders which can be reconstituted into solutions by addinga diluent before administration. Specifically excluded from the scope ofthe term “pharmaceutical formulation” are compositions for topicaladministration to patients, compositions for oral ingestion, andcompositions for parenteral feeding.

“Shelf life”, as used herein, means that the storage period during whichan active ingredient such as a therapeutic protein in a pharmaceuticalformulation has minimal degradation (e.g., not more than about 2-3%degradation) when the pharmaceutical formulation is stored underspecified storage conditions, for example, 2-8° C. Techniques forassessing degradation vary depending upon the identity of the protein inthe pharmaceutical formulation. Exemplary techniques includesize-exclusion chromatography (SEC)-HPLC to detect, e.g., aggregation,reverse phase (RP)-HPLC to detect, e.g., protein fragmentation, ionexchange-HPLC to detect, e.g., changes in the charge of the protein,mass spectrometry, fluorescence spectroscopy, circular dichroism (CD)spectroscopy, Fourier transform infrared spectroscopy (FT-IR), and Ramanspectroscopy to detect protein conformational changes. All of thesetechniques can be used singly or in combination to assess thedegradation of the protein in the pharmaceutical formulation anddetermine the shelf life of that formulation. Pharmaceuticalformulations preferably exhibit not more than about 2 to about 3%increases in degradation (e.g., fragmentation, aggregation or unfolding)over two years when stored at 2-8° C.

As used herein, “stable” formulations of biologically active proteinsare formulations that exhibit reduced aggregation and/or reduced loss ofbiological activity of at least 5% upon storage at 2-8° C. for at least2 years compared with a control formula sample, or alternatively whichexhibit reduced aggregation and/or reduced loss of biological activityunder conditions of thermal stress, e.g., 52° C. for 7-8 days.

As used herein, “viscosity” is a fluid's resistance to flow, and can bemeasured in units of centipoise (cP) or milliPascal-second (mPa-s),where 1 cP=I mPa-s, at a given shear rate. Viscosity can be measured byusing a viscometer, e.g., Brookfield Engineering Dial ReadingViscometer, model LVT (AMETEK Brookfield, Middleboro, Mass.). Viscositycan be measured using any other methods and in any other units known inthe art (e.g., absolute, kinematic or dynamic viscosity), understandingthat it is the percent reduction in viscosity afforded by use of theexcipients that is important. Regardless of the method used to determineviscosity, the percent reduction in viscosity in excipient formulationsversus control formulations will remain approximately the same at agiven shear rate.

As used herein, a formulation containing an amount of an excipienteffective to “reduce viscosity” (or a “viscosity-reducing” amount orconcentration of such excipient) means that the viscosity of theformulation in its final form for administration (if a solution, or if apowder, upon reconstitution with the intended amount of diluent) is atleast 5% less than the viscosity of an appropriate control formulation,such as those, for example, exemplified herein. Excipient-free controlformulations might also be used but may not always be the mostappropriate control formulation because such a formulation may not beimplementable as a therapeutic formulation due to hypotonicity, forinstance.

Similarly, a “reduced viscosity” formulation is a formulation thatexhibits reduced viscosity compared to a control formulation.

“High viscosity” describes the viscosity of, for example, apharmaceutical formulation that is difficult to handle, such as inmanufacturing or packaging, and/or administering the pharmaceuticalformulation to a subject (which can be device-dependent; for example,some devices are rated to deliver formulations having a viscosity of 8cP or less, thus any viscosity above 8 cP would be considered to have ahigh viscosity for that particular application, while other devices canbe rated to deliver formulations having a viscosity of 50 cP or less).In general, pharmaceutical formulations having a viscosity of at leastabout 10 cP, including for example, aleast about, in cP, 10, 11, 12, 14,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49,50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67,68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85,86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, and 100 or evengreater.

Protein therapeutics often need to be given at high doses, but forinjection, a smaller volume is often necessary which can result inincreased viscosity of the solution because the protein therapeuticconcentration is increased. When large doses of therapeutic protein areto be administered in a small volume of liquid (such as for injection),it is highly desirable to provide formulations with high concentrationsof protein that do not exhibit the increased viscosity typically seenwith such high protein concentrations.

High viscosity formulations are difficult to handle duringmanufacturing, including at the bulk and filling stages. High viscosityformulations are also difficult to draw into a syringe and inject, oftennecessitating use of lower gauge needles which can be unpleasant for thepatient. The addition of NMP to solutions of biologically active proteinunexpectedly reduced the viscosity of high concentration proteinsolutions.

“N-acetyl arginine” (NAR) means the molecule of formula 1.

The use of NMP permits a higher concentration of therapeutic proteins tobe used in the formulation without a concomitant increase in viscosity.Thus, provided herein are methods for stabilizing or reducing viscosityof protein formulations by adding NMP in an amount effective to reduceviscosity. Provided herein are also reduced viscosity formulations oftherapeutic proteins, including antibodies, containing effective amountsor concentrations of NMP. Also provided herein are methods of screeningone or more formulations, each containing different concentrations ofNMP to identify suitable or optimal concentrations that reduceviscosity. Further provided are methods of preparing a lyophilizedpowder from reduced-viscosity solution formulations described herein,and methods of reconstituting the lyophilized powders described hereinvia addition of a sterile diluent. In any of these aspects, additionalexcipients can be included, such as arginine, lysine, NAR, and mixturesthereof.

Thus provided herein are pharmaceutical formulations containingbiologically active polypeptides and viscosity-reducing concentrationsof NMP. Furthermore, such pharmaceutical formulations can furthercomprise additional excipients, such as arginine, lysine, NAR, andmixtures thereof. The reduction in viscosity is at least about 5%, suchas about at least 5%-90% versus control formulations. In one embodimentthe reduction in viscosity ranges from about 10-80%. In other exemplaryembodiments, the reduction in viscosity is at least 5%, at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 55%, at least 60%, atleast 65%, at least 70%, at least 75%, at least 80% or at least 85%.

Formulations can optionally include pharmaceutically acceptable salts,buffers, surfactants, other excipients, carriers, diluents, and/or otherformulation agents.

Exemplary pharmaceutically acceptable buffers include acetate (e.g.,sodium acetate), succinate (such as sodium succinate), glutamic acid,glutamate, gluconate or other organic acid buffers. Exemplary bufferconcentration can be from about 1 mM to about 100 mM, or from about 5 mMto about 50 mM, depending, for example, on the buffer and the desiredtonicity (e.g., isotonic, hypertonic or hypotonic) of the formulation.Exemplary pH's include from about 4.0 to about 6.0, or from about 4.8 toabout 5.5, or about 5.0, or greater than about 4.5, greater than about4.8, or greater than about 5.0, or greater than about 5.5.

Suitable diluents, other excipients, or carriers and other agentsinclude, but are not limited to, antioxidants, coloring, flavoring anddiluting agents, emulsifying agents, suspending agents, solvents,fillers, bulking agents, buffers, vehicles, diluents and/orpharmaceutical adjuvants. For example, a suitable vehicle can be,physiological saline solution, citrate buffered saline, or artificialCSF, possibly supplemented with other materials common in compositionsfor parenteral administration. Neutral buffered saline or saline mixedwith serum albumin are further exemplary vehicles. Those skilled in theart readily recognize a variety of buffers that can be used in thecompositions, and dosage forms disclosed herein. Typical buffersinclude, but are not limited to, pharmaceutically acceptable weak acids,weak bases, or mixtures thereof. Exemplary buffer components are watersoluble materials such as phosphoric acid, tartaric acids, lactic acid,succinic acid, citric acid, acetic acid, ascorbic acid, aspartic acid,glutamic acid, or salts thereof. Exemplary salts include inorganic andorganic acids, or bases such as metals or amines, in exemplaryconcentrations such as about 50-200 mM, or 100-200 mM, or about 100 mM,or about 150 mM.

Other excipients or stabilizers can also be included, for example,sugars (e.g., sucrose, glucose, trehalose, fructose, xylose, mannitose,fucose), polyols (e.g., glycerol, mannitol, sorbitol, glycol, inositol),amino acids or amino acid derivatives (e.g., arginine, proline,histindine, etc.) or surfactants (e.g., polysorbate, includingpolysorbate 20, or polysorbate 80, or poloxamer, including poloxamer188, or PLURONIC® F-68, or lecithin or protamine sulfate). Exemplaryconcentrations of surfactant can range to at least about 0.001%, suchas, for example, from about 0.001% to about 1.0%, or from about 0.002%to about 1.0%, or about 0.05 to 0.2%. Preservatives can also beincluded, such as benzyl alcohol, phenol, m-cresol, chlorobutanol orbenzethonium CI, e.g., at concentrations ranging from about 0.1% toabout 2%, or from about 0.5% to about 1%.

One or more other pharmaceutically acceptable carriers, excipients orstabilizers such as those described in Remington's PharmaceuticalSciences, 21st edition, Osol, A. Ed. (2005) can be included in theformulation provided that they do not adversely affect the desiredcharacteristics of the formulation.

The concentration of the therapeutic protein, such as an antibody, inthe formulation depends on the end use of the pharmaceutical formulationand can be easily determined by a person of skill in the art.

Therapeutic proteins that are antagonists are frequently administered athigher concentrations than those that are agonists. Particularlycontemplated high concentrations of therapeutic proteins (without takinginto account the weight of chemical modifications such as pegylation),including antibodies, are at least about 100, at least about 110, atleast about 120, at least about 130, at least about 140, at least about150, at least about 175, at least about 180, at least about 185, atleast about 190, at least about 195, at least about 200, at least about210, at least about 220, at least about 230, at least about 240, atleast about 250, or at least about 300 mg/mL, and/or less than about200, less than about 210, less than about 220, less than about 230, lessthan about 240, or less than about 250 mg/mL. Exemplary highconcentrations of therapeutic proteins, such as antibodies, in theformulation can range from at least about 100 mg/mL to about 1000 mg/mL.Additionally, high concentrations of therapeutic proteins, such asantibodies, in the formulations can range from at least about 100 mg/mLto about 300 mg/mL. Other protein concentrations (without taking intoaccount the weight of chemical modifications such as PEGylation), arealso contemplated, e.g., at least about 1, at least about 5, at leastabout 10, at least about 20, at least about 30, at least about 35, atleast about 40, at least about 45, at least about 50, at least about 55,at least about 60, at least about 65 or at least about 70 mg/mL.Disclosed herein are formulations and methods in which the concentrationof therapeutic protein results in a viscosity of at least 6, at leastabout 8, at least about 10, at least about 12, at least about 14, atleast about 16, at least about 18, at least about 20, at least about 25,at least about 30, at least about 35 cP or higher and the inclusion ofNMP results in the reduction of the viscosity by about 5% or greater.For example, a solution with a viscosity of about 20 cP may be difficultto inject with a standard 27 gauge needle. All references to mg/mLconcentration of therapeutic protein, weight of therapeutic protein (mg)or molecular weight of therapeutic protein (kD) herein mean therespective weight of the proteinaceous part of the therapeutic protein,excluding any non-proteinaceous modifications.

Disclosed herein are methods of reducing the viscosity of and/orimproving stability of a liquid pharmaceutical formulation of atherapeutic protein, by combining the therapeutic protein and aviscosity-reducing amount of NMP.

In exemplary embodiments, the therapeutic protein is at a high proteinconcentration as described above. In some embodiments, the reduction inviscosity is at least about 5%, at least about 10%, at least about 15%,at least about 20%, at least about 25%, at least about 30%, at leastabout 35%, at least about 40%, at least about 45%, at least about 50%,at least about 55%, at least about 60%, at least about 65%, at leastabout 70%, at least about 75%, at least about 80% or at least about 85%compared to control formulations

In another aspect, provided herein are liquid solutions comprising atherapeutic protein and NMP wherein the formulations exhibit reducedviscosity relative to control formulations. In exemplary embodiments,the therapeutic protein is at a high protein concentration as describedabove. In some embodiments, NMP is present at a viscosity-reducing(weight:volume) concentration. NMP can be used at concentrations up toits solubility limit. Such solutions can further comprise a sugar orother polyol such as sucrose or sorbitol, in an amount effective tofurther improve stability, reduce aggregation, and/or make theformulation isotonic, without significantly increasing viscosity.

In exemplary embodiments, the concentration of NMP is at least about 50mM to about 700 mM, or at least about 100 mM to about 450 mM, or atleast about 300 mM to about 425 mM. In exemplary embodiments theconcentration of NMP is at least about 50, at least about 100, at leastabout 150, at least about 200, at least about 250, at least about 300,at least about 350, at least about 400, or at least about 425 mM. Otherexemplary embodiments include concentrations of excipients effective tomake the formulation isotonic, without significantly increasingviscosity. Exemplary concentrations include those at least about 100 mMor greater, in further embodiments the amounts are at least about 300 mMor greater, in still further embodiments the amounts are at least about425 mM or greater.

Furthermore, in exemplary embodiments wherein NMP and NAR are bothpresent, the concentration of NMP is at least about 50 mM to about 700mM, or at least about 100 mM to about 450 mM, or at least about 300 mMto about 425 mM. The concentration of NAR is at least about 90-200 mM,or at least about 100-190 mM, or at least about 110-180 nM, or at leastabout 120-170 mM, or at least about 130-160 mM, or at least about140-150 mM, such as 155 mM. In exemplary embodiments the concentrationof NMP is at least about 50, at least about 100, at least about 150, atleast about 200, at least about 250, at least about 300, at least about350, at least about 400, or at least about 425 mM. In exemplaryembodiments, the concentration of NAR is at least about 90 mM, or atleast about 100 mM, or at least about 110 mM, or at least about 120 mM,or at least about 130 mM, or at least about 140 mM, or at least about150 mM, or at least about 155 mM, or at least about 160 mM, or at leastabout 170 mM, or at least about 180 mM, or at least about 190 mM, or atleast about 200 mM. Other exemplary embodiments include concentrationsof excipients effective to make the formulation isotonic, withoutsignificantly increasing viscosity. Exemplary concentrations includethose at least about 100 mM or greater, in further embodiments theamounts are at least about 300 mM or greater, in still furtherembodiments the amounts are at least about 425 mM or greater.

In exemplary embodiments, NMP is present at a concentration of at leastabout 50 mM, such as, for example, 50 mM, about 60 mM, about 70 mM,about 80 mM, about 90 mM, about 100 mM, about 110 mM, about 120 mM,about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM,about 180 mM, about 190 mM, about 200 mM, about 250 mM, about 275 mM,about 300 mM, about 325 mM, about 350 mM, about 375 mM, about 400 mM,about 425 mM, about 450 mM, about 475 mM, about 500 mM, about 525 mM,about 550 mM, about 575 mM, about 600 mM, about 625 mM, about 650 mM,about 675 mM, or about 700 mM. Preferred ranges of NMP include, but arenot limited to, 50 mM to 600 mM, 50 mM to 150 mM, 50 mM to 200 mM, and370-600 mM.

In exemplary embodiments, the excipients are both NMP and NAR, whereinNMP has a concentration of at last about 50 mM, such as, for example,about 50 mM, about 60 mM, about 70 mM, about 80 mM, about 90 mM, about100 mM, about 110 mM, about 120 mM, about 130 mM, about 140 mM, about150 mM, about 160 mM, about 170 mM, about 180 mM, about 190 mM, about200 mM, about 250 mM, about 275 mM, about 300 mM, about 325 mM, about350 mM, about 375 mM, about 400 mM, about 425 mM, about 450 mM, about475 mM, about 500 mM, about 525 mM, about 550 mM, about 575 mM, about600 mM, about 625 mM, about 650 mM, about 675 mM, or about 700 mM. Insuch embodiments, the concentration of NAR is at least about 90 mM, suchas, for example, about 90 mM, about 100 mM, about 110 mM, about 120 mM,about 130 mM, about 140 mM, about 150 mM, about 155 mM, about 160 mM,about 170 mM, about 180 mM, about 190 mM, or about 200 mM. In oneexemplary embodiment, the concentration of NMP is 100 mM and NAR is 155mM.

In another aspect, provided herein are lyophilized protein formulationscomprising a therapeutic protein NMP, wherein upon reconstitution withthe recommended amount of diluent, the formulations exhibit reducedviscosity relative to control formulations. In exemplary embodiments,the therapeutic protein is at a high protein concentration as describedabove. In some embodiments, NMP is present at an amount effective toreduce viscosity upon reconstitution with diluent (weight:weightconcentration). Such formulations can further comprise a sugar or otherpolyol such as sucrose or sorbitol, in an amount effective to furtherimprove stability, reduce aggregation, and/or make the formulationisotonic, without significantly increasing viscosity.

In exemplary embodiments of these formulations, the concentration of NMPis at least about 1 μg/mg therapeutic protein, up to about 1.0 mg/mgtherapeutic protein. In some embodiments, the concentration of NMP is atleast about 1, 10, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500 or550 μg/mg therapeutic protein. In other exemplary embodiments, theconcentration of NMP is up to about 600, 650, 700, 750, 800, 850, 900,950 or 1000 μg/mg therapeutic protein.

Provided herein are methods for stabilizing a liquid pharmaceuticalformulation comprising an antibody or fragment thereof at aconcentration of about 100 mg/mL to about 1000 mg/mL, such as, forexample, about 100 mg/L to about 300 mg/mL (e.g., about 100 mg/mL, about110 mg/mL, about 120 mg/mL, about 130 mg/mL, about 140 mg/mL, about 150mg/mL, about 160 mg/mL, about 170 mg/mL, about 180 mg/mL, about 190mg/mL, about 200 mg/mL, about 210 mg/mL, about 220 mg/mL, about 230mg/mL, about 240 mg/mL, about 250 mg/mL, about 260 mg/mL, about 270mg/mL, about 280 mg/mL, about 290 mg/mL, about 300 mg/mL). In exemplaryaspects, the liquid pharmaceutical formulation comprises N-methylpyrrolidone and the method comprises the step of combining the reducedviscosity liquid pharmaceutical formulation with a stabilizer selectedfrom the group consisting of polysorbate 20, polysorbate 80, PLURONIC®F-68, lecithin, and protamine sulfate. In exemplary aspects, the liquidpharmaceutical formulation comprises a combination of excipientscomprising N-methyl pyrrolidone and N-acetyl arginine and the methodcomprises the step of combining the reduced viscosity liquidpharmaceutical formulation with a stabilizer selected from the groupconsisting of polysorbate 20, polysorbate 80, PLURONIC® F-68, lecithin,and protamine sulfate. Accordingly, provided herein are methods forstabilizing a liquid pharmaceutical formulation comprising an antibodyor fragment thereof at a concentration of at least 140 mg/mL andN-methyl pyrrolidone. In exemplary aspects, the method comprises thestep of combining the reduced viscosity liquid pharmaceuticalformulation with a stabilizer selected from the group consisting ofpolysorbate 20, polysorbate 80, PLURONIC® F-68, lecithin, and protaminesulfate. Also, provided herein are methods for stabilizing a liquidpharmaceutical formulation comprising an antibody or fragment thereof ata concentration of at least 140 mg/mL and a combination of excipientscomprising N-methyl pyrrolidone and N-acetyl arginine. In exemplaryaspects, the method comprises the step of combining the reducedviscosity liquid pharmaceutical formulation with a stabilizer selectedfrom the group consisting of polysorbate 20, polysorbate 80, PLURONIC®F-68, lecithin, and protamine sulfate.

The term “stabilizer” as used herein refers to any agent that helpspharmaceutical formulations maintain the desirable properties of theproduct (e.g., therapeutic protein) until it is consumed by theend-user. In exemplary aspects, the stabilizer aids in the prevention ofprotein aggregate formation and/or of protein degradation products. Inexemplary aspects, the stabilizer is any agent that permits theformulation to exhibit a shelf-life of a desired length of time at adesired temperature. In exemplary aspects, the stabilizer is selectedfrom of polysorbate 20, polysorbate 80, PLURONIC® F-68, lecithin, andprotamine sulfate.

In exemplary aspects, the method stabilizes the liquid pharmaceuticalformulation such that little to no changes in stability is observedafter storage for a given time period at a given temperature, e.g., afreezing temperature, a refrigeration temperature, room temperature. Inexemplary aspects, the method stabilizes the liquid pharmaceuticalformulation such that little to no changes in stability is observedafter storage for a given time period at a temperature within about −50°C. to about 30° C., optionally, about −30° C., 2° C. to about 8° C., orabout 25° C. In exemplary instances, the method stabilizes the liquidpharmaceutical formulation such that little to no changes in stabilityis observed after storage at one of the above temperatures for a timeperiod of at least about 1 month, at least about 2 months, at leastabout 3 months, at least about 4 months, at least about 5 months, or atleast about 6 months. In exemplary instances, the method stabilizes theliquid pharmaceutical formulation such that little to no changes instability is observed after storage at one of the above temperatures fora time period greater than about 6 months, e.g., about 7 months orlonger, about 8 months or longer, about 9 months or longer, about 10months or longer, about 11 months or longer, about 12 months or longer.In exemplary instances, the method stabilizes the liquid pharmaceuticalformulation such that little to no changes in stability is observedafter storage at one of the above temperatures for a time period of atleast 1 year. In exemplary aspects, the methods stabilize the liquidpharmaceutical formulation to withstand one or more stress conditions,e.g., elevated temperature conditions, transportation conditions (e.g.,intermittent periods of agitation), shear stress, freeze-thawconditions, and the like. Stress conditions are described herein at,e.g., Example 3.

Stability can be assessed in many ways, including monitoringconformational change over a range of temperatures (thermostability)and/or time periods (shelf-life) and/or after exposure to stressfulhandling situations (e.g., physical shaking). Stability of formulationscontaining varying concentrations of formulation components can bemeasured using a variety of methods. For example, the amount of proteinaggregation can be measured by visual observation of turbidity, bymeasuring absorbance at a specific wavelength, by size exclusionchromatography (in which aggregates of a protein will elute in differentfractions compared to the protein in its native active state), HPLC, orother chromatographic methods. Other methods of measuring conformationalchange can be used, including using differential scanning calorimetry(DSC), e.g., to determine the temperature of denaturation, or circulardichroism (CD), which measures the molar ellipticity of the protein.Fluorescence can also be used to analyze the composition. Fluorescenceencompasses the release or absorption of energy in the form of light orheat, and changes in the polar properties of light. Fluorescenceemission can be intrinsic to a protein or can be due to a fluorescencereporter molecule. For example, 8-Anilinonaphthalene-1-sulfonic acid ANSis a fluorescent probe that binds to the hydrophobic pockets ofpartially unfolded proteins. As the concentration of unfolded proteinincreases, the number of hydrophobic pockets increases and subsequentlythe concentration of ANS that can bind increases. This increase in ANSbinding can be monitored by detection of the fluorescence signal of aprotein sample. Other means for measuring stability can be used and arewell known to persons of skill in the art.

In exemplary aspects, upon carrying out the methods provided herein, theliquid pharmaceutical formulation is stabilized, such that storage fortime periods and/or at temperature and/or under stress conditions, asdescribed above, alters the amount of protein degradation or the amountof protein aggregation by not more than about 15%, not more than about10%, or not more than about 5%, relative to the amounts before storage.

In yet another embodiment, provided herein are methods of preventingself-association of proteins in liquid formulations by using NMP as anexcipient in any of the amounts or concentrations described herein.Furthermore, such NMP-containing liquid formulations can furthercomprise arginine, lysine, NAR, and mixtures thereof. Formulations withimproved stability (e.g., reduced aggregation) and shelf-life are alsoprovided.

Also provided herein are kits comprising a liquid protein formulation asprovided herein, and instructions for its administration, optionallywith a container, syringe and/or other administration device. Furtherprovided herein are kits comprising a lyophilized protein formulation asprovided herein, optionally in a container, and instructions for itsreconstitution and administration, optionally with a vial of sterilediluent, and optionally with a syringe or other administration device.Exemplary containers include vials, tubes, bottles, single ormulti-chambered pre-filled syringes, or cartridges. Exemplaryadministration devices include syringes, with or without needles,infusion pumps, jet injectors, pen devices, transdermal injectors, orother needle-free injector, or an aerosolization device for nasal orpulmonary delivery.

In another aspect, a method is provided for screening for aviscosity-reducing concentration of NMP comprising the steps of: (1)assessing the viscosity of a first solution comprising a firstconcentration of NMP and a therapeutic protein, such as an antibody, (2)assessing the viscosity of a second solution comprising a differentsecond concentration of NMP and the therapeutic protein, and (3)determining that the first concentration of NMP is moreviscosity-reducing than the second concentration of NMP if the firstsolution is less viscous. Viscosity can be determined, e.g., using anAries ARG2 Rheometer or a Brookfield RV-DVIII Rheometer which isstabilized at 25° C. with a circulating temperature bath. Five hundredmicroliters of sample is pipetted into the rheometer and the rpmadjusted for percentage torque values between 10-80%. The samples areallowed to stabilize at that range and data points are collected.

Similar methods are provided for screening for an aggregation-reducingor stabilizing concentration of NMP.

The following examples are to aid the understanding of the claimedsubject matter, but are not to be construed as limiting the scope of theclaimed subject matter. All citations throughout the disclosure arehereby expressly incorporated by reference.

EXAMPLES Example 1 Materials and Methods Materials

Evolocumab (IgG2) concentrations varied from 120 to 225 mg/mL. Proteinconcentrations were determined by absorbance at 280 nm using anextinction coefficient of 1.48 M⁻¹ cm for evolocumab. Evolocumab drugsubstance was formulated in 10 mM acetate, pH5.2, 220 mM proline, 0.08%polysorbate 80.

Amino acids were reagent grade and purchased from Sigma Aldrich (St.Louis, Mo.). β-mercaptoethanol (β-ME), N-ethylmaleimide (NEM),dithiothreitol (DTT), iodoacetamide (IAA), calcium chloride and sodiumacetate were acquired from Sigma-Aldrich (St. Louis, Mo.). GMP N-Methylpyrrolidone was acquired from Ashland (Covington, Ky.). Glacial aceticacid, acetonitrile, trifluoroacetic acid and hydrochloric acid wereacquired from JT Baker (Center Valley, Pa.). The sodium dodecyl sulfatemolecular weight (SDS-MW) analysis kit was acquired from Beckman Coulter(Brea, Calif.). Trypsin of sequencing grade was purchased from Promega(Madison, Wis.) and stored at −20° C.

HYPAK SCF™ sterile 2.25 mL syringes and stoppers were obtained from(Becton Dickinson and Company (BD; Franklin Lakes, N.J.).

Preparation of Stability Samples and Ultrafiltration (UF)/Diafiltration(DF) Methodology

Drug substance was buffer exchanged into formulation buffer with a benchscale Millipore TFF UF/DF system using a Millipore PELLICON® XL filter,50 cm² size (regenerated cellulose, 30,000 molecular weight cut-off)membrane (EMD Millipore, Billerica, Mass.). The diafiltration step wasperformed until at least eight volumes of diafiltration buffer wereexchanged. Once the diafiltration step was completed, the UF/DF systemwas switched to ultrafiltration mode and each formulation wasconcentrated to the target concentration levels.

Formulations were buffered with 10 mM acetate, pH 5.0. NMP was the mainexcipient used in the study and its concentrations varied from 70-900mM. Post dialysis, polysorbate 80 or PLURONIC® F-68 was added toformulations as dictated by screening. Evolcumab stability samples wereincubated at the controlled temperatures of −30° C., 4° C., 25° C. and37° C. Incubation time varied between experiments.

Freeze-Thaw Cycles

Large scale freeze-thaw was performed immediately after UF DF proteinpreparation. Freeze-thaw was carried out in 250 mL polypropylenecontainers. Protein volume varied between 125-200 mL. Each 24 hourfreeze cycle was followed by room temperature thaw. Three cycles werecarried out prior to material fill into 3 cc vials.

Viscosity

Viscosity was measured using m-VROC™ rheometer (San Ramon, Calif.) witha circulating water bath at constant 25° C. A 500 μL glass syringe wasloaded with approximately 450 μL of sample and was locked into place forextrusion at shear rate of 1000/s. The viscosity reading was generatedby RheoSense software (RhoeSense, San Ramon, Calif.). Measurements areshown in centipoise and are an average of 3 true replicate readingsresulting from three independent readings.

Confirmatory readings were performed over a range of 1-1000/s, a MCR300rheometer (Anton PAAR, Ashland, Va.) equipped with a CP50-1 cone/platemeasuring system. Approximately 0.6 mL of samples were loaded onto thelower measuring plate and allowed to come to thermal equilibrium at 25°C. A solvent trap was used to prevent solvent evaporation. 700 s⁻¹ to1000 s⁻¹ values were averaged and reported for each sample measured dueto viscosity being relatively independent of shear rates in this range.

It was determined that the two rheometers generated comparable datasets, a finding that has been previously reported by others. Due to easeof use and lower volume requirements, most data was generated using them-VROC™ rheometer.

Concentration

A 40 μL aliquot of each sample was used to measure protein concentrationvia SoloVPE instrument (C Technologies Inc., Bridgewater, N.Y.)interfaced with a Varian CARY® 50 Bio UV/Visible Spectrometer (VarianInc., Palo Alto, Calif.). Sample dilution was not necessary with thisinstrumental method where even high protein concentration samples can bemeasured directly. Measurements are shown in mg/mL.

Size-Exclusion HPLC (SE-HPLC)

Evolocumab analysis was performed using a G3000SWxl Column, 7.8 mm ID×30cm, 5 μm particle and 250 Å pore column (Tosoh Corp, Grove City, Ohio).Isocratic elution with 100 mM sodium phosphate, 250 mM NaCl, pH 6.9 wasperformed at a flow rate of 0.5 mL/min for 37 minutes. Autosamplertemperature was set to 4° C. Column temperature was maintained at 25° C.Protein elution was monitored at 280 nm and occurred at a retention timeof approximately 16 minutes.

Reduced and Non-Reduced CE-SDS

Reduced capillary electrophoresis with sodium dodecyl sulfate (rCE-SDS)was performed by incubating samples in a 0.5% SDS, 50 mM Tris, pH 9.0buffer with 0.5% β-ME for 10 minutes at 70° C. Following incubation,samples were centrifuged at 10,000 rpm for 2 minutes and injected onto a30-cm bare fused silica capillary with a 20-cm effective length and a 50μm inner diameter using electrokinetic injection. Separation wasperformed using CE-SDS gel (Beckman Coulter, Brea, Calif.) and 15 kVeffective voltage. Detection was performed at 220 nm by UV absorbance.

Non-reducing solution was prepared by mixing 100 μL 140 mM NEM with1,900 μL non-reducing sample buffer composed of 10-60 mM sodiumphosphate, 2% SDS, pH 6.5. Larger volumes of non-reducing sample bufferwere generated by keeping the ratio of reagents constant. Incubation wasset at 60° C. for 5 minutes. Final concentrations in the incubatedsample were protein, 1.0 mg/mL; NEM, 4.7 mM; and SDS, 1.3%.

CEX

Evolocumab analysis was performed using a ProPac™ WCX-10, 4×250 mm(Thermo Scientific, Waltham, Mass.). Sample elution was performed at aflow rate of 0.5 mL/min. Autosampler temperature was set to 4° C. Columntemperature was maintained at 30° C. Mobile phase A was composed of 20mM MES, at pH 6.0±0.05; mobile phase B was composed of 20 mM MES, 0.5MNaCl at pH 6.0±0.05. Sample was loaded to a pre-equilibrated column at0% mobile phase B. The ramp of 0-30% took place over a period of 20minutes, followed by 8 minute wash step at 100% mobile phase B andre-equilibration of the column to initial conditions for a period of 8minutes. Protein elution was monitored at 280 nm and occurred at aretention time of approximately 23 minutes.

Trypsin Mapping Reduction, Alkylation and Trypsin Digestion:

Approximately 1 mg of evolocumab was denatured with 6 M guanidine HCl,0.25 M Tris-HCl, 1 mM EDTA, at pH 7.5. Twenty microliters of 0.5 Mdithiothreitol (DTT) was added to the solution and the reaction mixturewas placed at 37° C. for 40 min. After the sample was cooled to roomtemperature, 24 μl of 0.5 M iodoacetamide (IAA) was added and the samplewas incubated at room temperature in the dark for 30 min. Tenmicroliters of 0.5 M DTT was added to the samples in order to terminatethe alkylation reaction. Approximately 500 μL of reduced and alkylatedmaterial was buffer exchanged with 750 μl of 0.1 M Tris-HCl to a finalconcentration of 1 mg/mL of evolocumab using a NAP™-5 column (AmershamBioSciences, Uppsala, Sweden) equilibrated with 10 mL of 50 mM Tris, 1mM CaCl₂, pH 7.0. Trypsin digestion was performed for 4.5 h at 37° C.using an enzyme:protein ratio of 1:55. The digests were thenrefrigerated at 4° C. for analysis.

HPLC Separation of Digested Peptides and Mass Spectrometry (MS)Detection and Analysis.

The reduced evolocumab was analyzed using an Agilent 1100 HPLC unitequipped with a diode-array detector, autosampler, micro flow cell andtemperature controlled column compartment (Agilent, Palo Alto, Calif.,USA). Autosampler temperature was set to 4° C. Column was heated at 45°C. to enhance separation. Mobile phase A consisted of 0.1%trifluoroacetic acid in water and mobile phase B consisted of 95%acetonitrile, 5% H₂O and 0.1% trifluoroacetic acid. Separation wasperformed on a Varian Polaris™ C18-A 5 micron column (PN A2000-250X020).The column was equilibrated at 2% solvent B. Two minutes after sampleinjection the concentration of buffer B was increased to 22% over 38 minfollowed by a linear gradient of 22% to 42% B over 100 min. The columnwas re-equilibrated by ramping up buffer B to 100% over 5 min, 5 min at100% B, dropping down to 2% B over 1 min followed by 15 min at 2% B. UVabsorption was monitored at 214 nm.

Mass spectrometry was performed on a Bruker ESI-MSD. The ESI-MSD massspectrometer was set to run in positive ion mode with a nebulizerpressure of 40 psi, dry gas flow of 8 L/min, dry gas temperature of 350°C., and m/z range of 50-2200 m/z. The deconvolution of electrosprayionization mass spectra was performed using QuantAnalysis.

NMP Quantitation Method

Samples were separated using Luna C8 column (150×2.1 mm, 5 pm particlesize, 100 A, Phenomenex, Torrance, Calif.). Mobile phase A was 0.05% TFAin water, mobile phase B was acetonitrile. The gradient profile began byholding the mobile phase composition for 8 min at 85:15 (A:B), followedby a linear gradient to 15:85 for over 1 min. The composition was heldat 15:85 for 5 min, then returned in 1 min to 85:15 and equilibrated forat least 8 min before the next injection. The total mobile phase flowrate was 1.0 mL/min, and both mobile phases were filtered and degassedbefore use. Column temperature was maintained at 40° C. The injectionvolume was 100 μL for both samples and standards. The detectorwavelength was 220 nm. Samples and standards were diluted prior toinjection. Standard curves were generated for each set of unknownstested within a sequence.

Example 2 Viscosity Profiles Using Organic Solvents

A formulation of evolocumab that utilized isotonic levels of proline wascompared against a number of alternate formulations containing arginineand polar solvents DMA, DMSO, and NMP. As shown in FIG. 1A, the argininecontaining formulation was significantly less viscous than proline basedformulation. At approximately 1000 mM levels DMA, DMSO and NMP alldecreased evolocumab viscosities more effectively than amino acidformulations. Among the polar solvents tested, NMP was most effective inlowering evolocumab viscosity. While 1000 mM polar solventconcentrations resulted in highly hyperosmotic solutions, 340 mM NMPresulted in a formulation that is isotonic and still marks a distinctlydecreased viscosity profile of evolocumab. Titration studies showed thatthe viscosity of evolocumab decreased sharply with increasingconcentrations of NMP up to 700 mM; above 700 mM the viscosity ofevolocumab appeared to reach a plateau and was nearly independentfurther addition of solvent (FIG. 1B).

VP-DSC experiments showed that increasing levels of NMP in solutioncause a decrease in protein unfolding temperature (Tm) for both domainsof evolocumab. No notable changes in the enthalpy of unfolding (delta H)were noted with increase of NMP in solution. Thus, while NMP is aparenterally approved excipient, lowering levels of NMP in theformulation can be advantageous as it is likely to result in a morestable protein profile. Whihle there is an indication that polarsolvents reduce solution viscosity through hydrophobic solvation (T. J.Kamerzell, et al., Pharmaceutical Biotechnology, 102:1182-1193 (2013)),arginine has been suggested to decrease viscosity by weakeningintra-protein electrostatic interactions (N. Inoue, et al. Journal ofBioscience and Bioengineering. 117(5):539-543 (2014)). It wascontemplated that a combination of excipients that work throughdifferent molecular mechanisms might result in still further viscosityreduction. FIG. 2 shows that a combination arginine and NMP leads toevolocumab viscosity curve that is very similar to formulationcontaining higher levels of NMP as a sole excipient. Further advantagesof having this combination formulation can be a potentially improvedstability profile due to presence of arginine (S. Yadav et al., J.Pharm. Sci. 99:1152-1168 (2010); T. Arakawa, et al., Biophys. Chem. 127:1-8 (2007); H. Hamada, et al., Curr. Pharm. Biotechnol. 10: 400-407(2009); M. Ishibashi, et al., Protein Expr. Purif. 42:1-6 (2005); K.Tsumoto, et al., Protein Pept. Lett. 12: 6130-619 (2005))).

Example 3 Antibody Stability in NMP-Based Formulations

While viscosity lowering effects are paramount to a design of a highconcentration protein formulation, protein stability must also beconsidered. To address this issue, the effects of NMP was evaluated onevolocumab quiescent storage stability in liquid and frozen state,stability during transport simulation and large scale freeze thaw cyclesas well as for potential interactions with pre-filled syringes.

Evolocumab was originally formulated in 10 mM acetate, pH 5.0, 340 mMNMP, and 0.01% polysorbate 80 at 210 mg/mL. Under these conditions,evolocumab showed increased levels of aggregation at elevatedtemperatures. At 210 mg/mL evolocumab aggregation rates at 25 C in NMPformulation were 10 times higher than those observed at 140 mg/mLevolocumab in proline based formulation. While some of this is likelydue to increased protein concentration, NMP was contributing toincreased aggregation rates. To mitigate the high aggregation ratesobserved under these conditions, formulation screening was performed.The additional excipient screening was limited to parenterally approvedcompounds of high molecular weight as those have minimal additionalcontribution to solution osmolality. The compounds also needed to notraise the final solution viscosity.

PEG 3350, PEG4000, lethicin, protamine sulfate, and PLURONIC® F-68 wereevaluated. While lethicin and protamine sulfate showed improvedevolocumab stability and did not increase viscosity, these compoundsexhibited solubility issues upon freeze thaw cycles. PLURONIC® F-68 metall the selection criteria of improving evolocumab stability at elevatedtemperatures, having minimal contribution to osmolality and no negativeimpact on stability.

Evolocumab formulated in 10 mM acetate, pH 5.0, 340 mM NMP, 0.1%PLURONIC® F-68 was subjected to three cycles of large-scale freeze-thawat −30° C. followed by transportation simulation and long term storageat 2-8° C., at elevated temperatures and in frozen state. Concentrationof evolocumab to 210 mg/mL resulted in lower main peak by SEC however nochanges were noted in the initial levels of main peak by CEX whencompared with frozen control at 140 mg/mL.

Table 1 summarizes evolocumab stability after the above mentioned stressconditions in 3 cc glass vials. Upon completion of the freeze-thawcycles, evolocumab showed no change in stability profile by any of theassays. Transportation stress resulted in 0.3% main peak loss by SEC.After six months at 2-8° C. evolocumab showed no physical or chemicaldegradation, no particulation and retained full activity by potencyassay.

TABLE 1 Evolocumab liquid and frozen stability in 10 mM acetate, 340 mMNMP, 0.1% PLURONIC ® F-68, pH 5.0 Quiescent stability data is based on 6months of storage. Transportation Stability profile Stability profileStability profile Assay Large Scale FT stress (2-8° C.) (−30° C.) (25°C.) SEC no change 0.3% main peak no change 0.1% main peak 0.6% main peakloss loss/month loss/month WCX no change 0.8% main peak loss/monthPeptide map no change NA (multiple sites) rCE-SDS no change MFI nochange Visual no particulation Potency Full potency NA

TABLE 2 Comparison of liquid and frozen stability in NMP based and aminoacid-based formulations reported as percent per month losses 10 mMacetate, pH 5.0, 10 mM acetate, pH 5.0, 70 mM NMP, 130 mM 10 mM acetate,pH 5.0, 340 mM NMP, 0.1% arginine, 0.01% 220 mM proline, 0.01% pluronicF68 polysorbate 80 polysorbate 80 (210 mg/mL) (210 mg/mL) (140 mg/mL)Assay SEC CEX SEC CEX SEC CEX   37° C. 2.9 4.0 2.8 4.0 2.9 7.9   25° C.0.6 0.8 0.6 0.7 0.3 0.9    4° C. 0 0 0 0 0 0 −30° C. 0.1 0 0 0 0 0

Aggregation rates at 25° C. were higher than those observed in theproline based 140 mg/mL formulation (Tables 1 and 2). Concentrationdependent increase in aggregation rate is known to occur and is notunexpected. It is interesting to note that deamidation rates in NMPbased formulation were lower at 37° C. than those observed in prolineformulation under the same conditions (Table 2). This dynamic is likelydue deamidation rates being typically lower in more viscous samples aspreviously reported by Li et al (J. Phys. Chem. B. 117(21):6373-84(2013)).

Under −30° C. storage conditions, evolocumab showed 0.1% main peak lossper month by SEC but no chemical degradation, no participation andretention of full activity by potency assay. The data suggested that theNMP based formulation might be more suitable for liquid evolocumabstorage.

Evolocumab's stability profile formulated under the same conditions wasalso monitored during quiescent storage in 2.25 mL stacked needlepre-filled syringes. After 5 months of storage at 2-8° C., the stabilityprofile mimicked data reported for vial based storage.

A combination formulation using arginine and NMP (10 mM acetate, pH 5.0,130 mM arginine, 70 mM NMP, 0.01% polysorbate 80) was also evaluatedusing the same stress conditions as described above. Initial data showedno stability profile changes after large scale freeze-thaw cycles ortransportation stress. Furthermore after two months of storage under2-8° C. and −30° C. conditions, evolocumab showed no physical orchemical degradation, no participation and retained full activity bypotency assay. Aggregation rates at 25° C. and 37° C. were similar tothose observed in the 340 mM NMP formulation (Table 2). The datasuggests that a combination of arginine and NMP leads to improved frozenevolocumab stability as compared to the formulations that include NMPonly. Because arginine NMP formulation has a viscosity profile verysimilar to 340 mM NMP formulation, it is a more promising overallcandidate for evolocumab formulation.

Example 4 Viscosity Profile Using Lysine

FIG. 3 describes the behavior of evolocumab viscosity as a function ofthe concentration of a few different excipients investigated. Evolocumabin proline (“x”) could not be measured beyond 170 mg/mL due to gelationof the protein. The squares show the increase in viscosity of evolocumabas a function of protein concentration for a formulation containing 10mM sodium phosphate, 89 mM lysine at pH 6.2. At 192 mg/mL, the viscositywas >70 cP. This solution was divided into two parts. To one part,lysine was spiked in to achieve increments of 30 mM final concentration.To another part, buffer was added without lysine. It was seen that thebuffer arm (triangles) caused a viscosity decrease along the sameviscosity curve. However, when additional lysine was added, it droppedthe viscosity much more than just a dilution with buffer. Specifically,addition of lysine to go from 89 mM to ^(˜)120 mM dropped the viscosityfrom ^(˜)75 cP to ^(˜)30 cP while with just addition of buffer withoutlysine the viscosity drop was ^(˜)75 cP to ^(˜)60 cP.

With this information, two formulations of NMP were prepared, that wentthrough typical manufacturing stresses of UF/DF, freeze/thaw andtransportation. One formulation had 340 mM NMP and PLURONIC® F-68,another had 100 mM NMP, 120 mM lysine and 0.01% polysorbate 80. Theseformulations, which were filled into 2.25 mL prefilled syringes, werecompared for their attributes with respect to the control evolocumabformulation as shown in Table 3. It was seen that the key performancemeasures of each formulation were all within specifications and similarto the control formulation. The purity by SEC was slightly lower for theNMP alone formulation, but further analyses showed that this slightdecrease in main peak by SEC was a function of the high proteinconcentration; upon dilution to 170 mg/mL, it went up to 98.5%. Thus theproduct quality attributes appeared to be acceptable after the relevantmanufacturing stresses were applied.

TABLE 3 Viscosity of evolocumab formulations Formulation 10 mM acetate10 mM acetate 10 mM acetate pH 5.0 pH 5.0 pH 5.0 100 mM NMP 220 mMproline 340 mM NMP 120 mM lysine-HCl 0.01% PS80 0.1% PLURONIC ® F-680.01% PS80 136 mg/mL Attribute 210 mg/mL 210 mg/mL (control) Purity bySEC 97.6% 98.5% 98.7% Spec: >97% main peak Purity by CEX 59.4% 60.8%59.9% Spec: >51% main peak Subvisible particle counts >10 micron: 1376 ±238 >10 micron: 1211 ± 340 >10 micron: 1399 ± 530 Spec: >10 μm: 6000 >25micron: 317 ± 113 >25 micron: 218 ± 103 >25 micron: 275 ± 216 particlesor less >25 μm: 600 particles or less Visual inspection Practically freeof particles Practically free of Practically free of Spec: practicallyfree of particles particles particles Viscosity 35.5 cP 45.5 cP <15 cPat 136 mg/mL >>70 cP at 210 mg/mL

Example 5 The Viscosity Lowering Effect of NMP is Not Limited toEvolocumab

The effect of addition of NMP to five additional monoclonal antibodieswas tested, as shown in FIG. 4. There were three IgG2 antibodies tested(in addition to evolocumab, which is an IgG2), and three IgG1 antibodiestested. The data showed that for all molecules where the viscosity washigh (>20 cP) at high protein concentration (>150 mg/mL), the additionof NMP significantly reduced the viscosity of that antibody,irrespective of whether it was IgG1 or IgG2. However, if the viscosityin the control was low (<15 cP) to begin with, then addition of NMP didnot have any significant impact as can be expected.

Example 6 Viscosity of Formulations Lacking a Buffer System

Evolocumab samples were buffer exchanged using CENTRICON® centrifugalunits (Millipore Sigma; Billerica, Mass.) into the desired formulationsof 10 mM acetate pH 5.0, 340 mM NMP, 0.1% PLURONIC® F-68 and 340 mM NMP,0.1% PLURONIC® F-68. The pH of post-buffer exchange samples was adjustedon buffer-free formulation from pH 5.5 to pH 5.0. Samples weresterile-filtered using a 0.2 μm filter prior to concentration andviscosity measurements. Final adjusted sample concentration wasdetermined with SoloVPE instrument as previously described. Viscosityreadings were measured with RheoSense mVROC™ as previously described,and readings were an average of three replicate measurements.

As shown in Table 4, the attributes of pH and viscosity were comparablefor evolocumab samples formulated with and without a buffer system.

TABLE 4 Viscosity of evolocumab formulations with and without a buffersystem Formulation 10 mM acetate pH 5.0 340 mM NMP 340 mM NMP 0.1%PLURONIC ® F-68 0.1% PLURONIC ® F68 Attribute 210 mg/mL evolocumab 210mg/mL evolocumab pH 5.0 5.0 Visual Practically free of particlesPractically free of particles inspection Spec: practically free ofparticles Viscosity 35.7 ± 1.5 36.5 ± 2.1

Example 7 Viscosity of Formulations Comprising Both NMP and NAR(Prophetic)

Therapeutic protein samples (such as those of evolocumab) are bufferexchanged using, for example, CENTRICON® centrifugal units (MilliporeSigma; Billerica, Mass.) into the desired formulation of 10 mM acetate,100 mM NMP, 150-155 mM NAR, 0.01% polysorbate 80, pH 5.0 as indicated inTable 4.

TABLE 4 Target and ranges component target range acetate  10 mM  5-20 mMNMP 100 mM 50-150 mM NAR 155 mM 90-200 mM polysorbate 80 0.01% 0.005-0.02% pH 5.0 4.5-5.5The samples are analyzed as described in Example 1, including measuringconcentrations, viscosity, and therapeutic protein stability.

1. A pharmaceutical formulation, comprising an antibody or an antigenbinding fragment thereof at a concentration greater than about 100mg/mL, and N-methyl pyrrolidone at a concentration of about 100 mM toabout 500 mM , wherein the pH of the formulation is between about 4.0 toabout 8.0.
 2. The pharmaceutical formulation of claim 1, furthercomprising a buffer.
 3. A pharmaceutical formulation, comprising anantibody or an antigen binding fragment thereof at a concentrationgreater than about 100 mg/mL, about 1 mM to about 1000 mM N-methylpyrrolidone and about 90 mM to about 200 mM N-acetyl arginine, whereinthe pH of the formulation is between about 4.0 to about 8.0.
 4. Thepharmaceutical formulation of claim 3, further comprising a buffer. 5.The pharmaceutical formulation of any one of claims 1-4, wherein theantibody is selected from the group consisting of abagovomab, abciximab,actoxumab, adalimumab, afelimomab, afutuzumab, alacizumab, alacizumabpegol, ald518, alemtuzumab, alemtuzumab, alirocumab, altinumab,altumomab, amatuximab, anatumomab mafenatox, anrukinzumab, apolizumab,arcitumomab, aselizumab, atlizumab, atorolimiumab, bapineuzumab,basiliximab, bavituximab, bectumomab, belimumab, benralizumab,bertilimumab, besilesomab, bevacizumab, bezlotoxumab, biciromab,bivatuzumab, bivatuzumab mertansine, blinatumomab, blosozumab,brentuximab vedotin, briakinumab, brodalumab, canakinumab, cantuzumabmertansine, cantuzumab mertansine, caplacizumab, capromab pendetide,carlumab, catumaxomab, cc49, cedelizumab, certolizumab pegol, cetuximab,citatuzumab bogatox, cixutumumab, clazakizumab, clenoliximab,clivatuzumab tetraxetan, conatumumab, cr6261, crenezumab, dacetuzumab,daclizumab, dalotuzumab, daratumumab, demcizumab, denosumab, detumomab,dorlimomab aritox, drozitumab, duligotumab, dupilumab, ecromeximab,eculizumab, edobacomab, edrecolomab, efalizumab, efungumab, elotuzumab,elsilimomab, enavatuzumab, enlimomab pegol, enokizumab, enokizumab,enoticumab, enoticumab, ensituximab, epitumomab cituxetan, epratuzumab,erenumab, erlizumab, ertumaxomab, etaracizumab, etrolizumab, evolocumab,exbivirumab, exbivirumab, fanolesomab, faralimomab, farletuzumab,fasinumab, fbta05, felvizumab, fezakinumab, ficlatuzumab, figitumumab,flanvotumab, fontolizumab, foralumab, foravirumab, fresolimumab,fulranumab, futuximab, galiximab, ganitumab, gantenerumab, gavilimomab,gemtuzumab ozogamicin, gevokizumab, girentuximab, glembatumumab vedotin,golimumab, gomiliximab, gs6624, ibalizumab, ibritumomab tiuxetan,icrucumab, igovomab, imciromab, i mgatuzumab, inclacumab, indatuximabravtansine, infliximab, inolimomab, inotuzumab ozogamicin, intetumumab,ipilimumab, iratumumab, itolizumab, ixekizumab, keliximab, labetuzumab,lebrikizumab, lemalesomab, lerdelimumab, lexatumumab, libivirumab,ligelizumab, lintuzumab, lirilumab, lorvotuzumab mertansine,lucatumumab, lumiliximab, mapatumumab, maslimomab, matuzumab,mavrilimumab, mepolizumab, metelimumab, milatuzumab, minretumomab,mitumomab, mogamulizumab, morolimumab, motavizumab, moxetumomabpasudotox, muromonab-cd3, nacolomab tafenatox, namilumab, naptumomabestafenatox, narnatumab, natalizumab, nebacumab, necitumumab,nerelimomab, nesvacumab, nimotuzumab, nivolumab, nofetumomab merpentan,ocaratuzumab, ocrelizumab, odulimomab, ofatumumab, olaratumab,olokizumab, omalizumab, onartuzumab, oportuzumab monatox, oregovomab,orticumab, otelixizumab, oxelumab, ozanezumab, ozoralizumab,pagibaximab, palivizumab, palivizumab, panitumumab, panobacumab,parsatuzumab, pascolizumab, pateclizumab, patritumab, pemtumomab,perakizumab, pertuzumab, pexelizumab, pidilizumab, pintumomab,placulumab, ponezumab, priliximab, pritumumab, PRO 140, quilizumab,racotumomab, radretumab, rafivirumab, ramucirumab, ranibizumab,raxibacumab, regavirumab, reslizumab, rilotumumab, rituximab,robatumumab, roledumab, romosozumab, rontalizumab, rovelizumab,ruplizumab, samalizumab, sarilumab, satumomab pendetide, secukinumab,sevirumab, sibrotuzumab, sifalimumab, siltuximab, simtuzumab,siplizumab, sirukumab, solanezumab, solitomab, sonepcizumab, sontuzumab,stamulumab, sulesomab, suvizumab, tabalumab, tacatuzumab tetraxetan,tadocizumab, talizumab, tanezumab, taplitumomab paptox, tefibazumab,tefibazumab, telimomab aritox, telimomab aritox, tenatumomab,tenatumomab, teneliximab, teplizumab, teprotumumab, TGN1412,ticilimumab, tigatuzumab, tildrakizumab, TNX-650, tocilizumab,tocilizumab, toralizumab, tositumomab, tralokinumab, trastuzumab,TRBS07, tregalizumab, tremelimumab, tremelimumab, tucotuzumabcelmoleukin, tuvirumab, ublituximab, urelumab, urtoxazumab, ustekinumab,vapaliximab, vatelizumab, vedolizumab, veltuzumab, vepalimomab,vesencumab, visilizumab, volociximab, vorsetuzumab mafodotin, votumumab,zalutumumab, zanolimumab, zatuximab, ziralimumab, zolimomab aritox andthose shown in Table A.
 6. The pharmaceutical formulation of claim 2 or4, wherein the antibody is an anti-PCSK9 antibody.
 7. The pharmaceuticalformulation of claim 6, wherein the antibody is evolocumab.
 8. Thepharmaceutical formulation of any one of claims 1-4, wherein theantibody or the antigen binding fragment thereof is present at aconcentration between about 100 mg/mL to about 300 mg/mL.
 9. Thepharmaceutical formulation of claim 8, wherein the antibody is presentat a concentration of between about 180 mg/mL to about 230 mg/mL. 10.The pharmaceutical formulation of any one of claims 1-4, wherein theN-methyl pyrrolidone is at a concentration about 100 mM to about 425 mM.11. The pharmaceutical formulation of claim 9, wherein the N-methylpyrrolidone is at a concentration ofabout 100 mM.
 12. The pharmaceuticalformulation of claim 9, wherein the N-methyl pyrrolidone is at aconcentration of about 200 mM.
 13. The pharmaceutical formulation ofclaim 9, wherein the N-methyl pyrrolidone is at a concentration of about340 mM.
 14. The pharmaceutical formulation of claim 9, wherein theN-methyl pyrrolidone is at a concentration of about 425 mM.
 15. Thepharmaceutical formulation of claim 1 or 2, further comprising arginine.16. The pharmaceutical formulation of claim 15, wherein the arginine isat a concentration of about 90 to about 160 mM.
 17. The pharmaceuticalformulation of claim 9, further comprising arginine, wherein thearginine is at a concentration of about 90 to about 160 mM.
 18. Thepharmaceutical formulation of claim 9, further comprising arginine,wherein the arginine is at a concentration of about 120 mM.
 19. Thepharmaceutical formulation of claim 1 or 2, further comprising lysine.20. The pharmaceutical formlulation of claim 19, wherein the lysine isat a concentration of about 100 to about 120 mM.
 21. The pharmaceuticalformulation of claim 9, further comprising lysine, wherein the lysine isat a concentration of about 100 to about 120 nM.
 22. The pharmaceuticalformulation of claim 11, further comprising lysine, wherein the lysineis at a concentration of about 100 to about 120 nM.
 23. Thepharmaceutical formulation of claim 12, further comprising lysinewherein the lysine is at a concentration of about 100 to about 120 nM.24. The pharmaceutical formulation of any one of claims 1-4, wherein thepH of the formulation is between about 4.8 and 5.2.
 25. Thepharmaceutical formulation of claim 11, wherein the pH of theformulation is between about 4.8 and 5.2.
 26. The pharmaceuticalformulation of claim 12, wherein the pH of the formulation is betweenabout 4.8 and 5.2.
 27. The pharmaceutical formulation of claim 13,wherein the pH of the formulation is between about 4.8 and 5.2.
 28. Thepharmaceutical formulation of claim 14, wherein the pH of theformulation is between about 4.8 and 5.2.
 29. The pharmaceuticalformulation of claim 21, wherein the pH of the formulation is betweenabout 4.8 and 5.2.
 30. The pharmaceutical formulation of any one ofclaims 1-4, further comprising a stabilizer selected from the groupconsisting of polysorbate 20, polysorbate 80, PLURONIC® F-68, lecithin,and protamine sulfate.
 31. The pharmaceutical formulation of claim 30,wherein the stabilizer is at a concentration of between about 0.005% toabout 1.0%.
 32. The pharmaceutical formulation of claim 31, wherein thestabilizer is polysorbate 80 and is at a concentration of about 0.01%.33. The pharmaceutical formulation of claim 31, wherein the stabilizeris PLURONIC® F-68 and is at a concentration of between about 0.05% toabout 0.2%.
 34. The pharmaceutical formulation of claim 31, wherein thestabilizer is protamine sulfate and is at a concentration of about0.12%.
 35. The pharmaceutical formulation of claim 24, furthercomprising a stabilizer selected from the group consisting ofpolysorbate 20, polysorbate 90, PLURONIC® F-68, lecithin, and protaminesulfate.
 36. The pharmaceutical formulation of claim 33, wherein thestabilizer is PLURONIC® F-68 and is at a concentration of between about0.05% to about 0.2%.
 37. The pharmaceutical formulation of claim 25,further comprising a stabilizer selected from the group consisting ofpolysorbate 20, polysorbate 80, PLURONIC® F-68, lecithin, and protaminesulfate.
 38. The pharmaceutical formulation of claim 35, wherein thestabilizer is PLURONIC® F-68 and is at a concentration of between about0.05% to about 0.2%.
 39. The pharmaceutical formulation of claim 28,further comprising a stabilizer selected from the group consisting ofpolysorbate 20, polysorbate 80, PLURONIC® F-68, lecithin, and protaminesulfate.
 40. The pharmaceutical formulation of claim 37, wherein thestabilizer is polysorbate 80 and is at a concentration of about 0.01%.41. The pharmaceutical formulation of claim 2 or 4, wherein the bufferis selected from the group consisting of acetate buffer, succinatebuffer and glutamate buffer.
 42. The pharmaceutical formulation of claim41, wherein the buffer is at a concentration of about 5 mM to about 50mM.
 43. The pharmaceutical formulation of claim 9 comprising a buffer,wherein the buffer is sodium acetate buffer and it is at a concentrationof about 10 mM.
 44. The pharmaceutical formulation of claim 10comprising a buffer, wherein the buffer is sodium acetate buffer and itis at a concentration of about 10 mM.
 45. The pharmaceutical formulationof claim 11 comprising a buffer, wherein the buffer is sodium acetatebuffer and it is at a concentration of about 10 mM.
 46. Thepharmaceutical formulation of claim 12 comprising a buffer, wherein thebuffer is sodium acetate buffer and it is at a concentration of about 10mM.
 47. The pharmaceutical formulation of claim 13 comprising a buffer,wherein the buffer is sodium acetate buffer and it is at a concentrationof about 10 mM.
 48. The pharmaceutical formulation of claim 14comprising a buffer, wherein the buffer is sodium acetate buffer and itis at a concentration of about 10 mM.
 49. The pharmaceutical formulationof claim 16 comprising a buffer, wherein the buffer is sodium acetatebuffer and it is at a concentration of about 10 mM.
 50. Thepharmaceutical formulation of claim 17 comprising a buffer, wherein thebuffer is sodium acetate buffer and it is at a concentration of about 10mM.
 51. The pharmaceutical formulation of claim 18 comprising a buffer,wherein the buffer is sodium acetate buffer and it is at a concentrationof about 10 mM.
 52. The pharmaceutical formulation of claim 20comprising a buffer, wherein the buffer is sodium acetate buffer and itis at a concentration of about 10 mM.
 53. The pharmaceutical formulationof claim 21 comprising a buffer, wherein the buffer is sodium acetatebuffer and it is at a concentration of about 10 mM.
 54. Thepharmaceutical formulation of claim 22 comprising a buffer, wherein thebuffer is sodium acetate buffer and it is at a concentration of about 10mM.
 55. The pharmaceutical formulation of claim 23 comprising a buffer,wherein the buffer is sodium acetate buffer and it is at a concentrationof about 10 mM.
 56. A pharmaceutical formulation comprising about 340 mMN-methyl pyrrolidone, about 0.1% PLURONIC® F-68, and about 210 mg/mLevolocumab.
 57. The pharmaceutical formulation of claim 56, furthercomprising a buffer.
 58. The pharmaceutical formulation of claim 57,wherein the buffer is acetate.
 59. The pharmaceutical formulation ofclaim 58, wherein the acetate is present at a concentration of about 10mM.
 60. The pharmaceutical formulation of any one of claims 56-59,having a pH of about 5.0.
 61. A pharmaceutical formulation comprisingabout 100 mM N-methyl pyrrolidone, about 120 mM lysine, 0.01%polysorbate 80, and about 210 mg/mL evolocumab.
 62. The pharmaceuticalformulation of claim 61, further comprising a buffer.
 63. Thepharmaceutical formulation of claim 62, wherein the buffer is acetate.64. The pharmaceutical formulation of claim 63, wherein the acetate ispresent at a concentration of about 10 mM.
 65. The pharmaceuticalformulation of any one of claims 61-63, having a pH of about 5.0.
 66. Apharmaceutical formulation comprising about 100 mM N-methyl pyrrolidone,about 150 mM to about 155 mM N-acetyl arginine, 0.01% polysorbate 80,and about 210 mg/mL evolocumab.
 67. The pharmaceutical formulation ofclaim 66, further comprising a buffer.
 68. The pharmaceuticalformulation of claim 67, wherein the buffer is acetate.
 69. Thepharmaceutical formulation of claim 68, wherein the acetate is presentat a concentration of about 10 mM.
 70. The pharmaceutical formulation ofany one of claims 66-69, having a pH of about 5.0.
 71. A method forreducing the viscosity of a liquid pharmaceutical formulation comprisingan antibody or fragment thereof at a concentration of at least 140mg/mL, comprising the step of combining the antibody or fragment thereofwith about 100 mM to about 500 mM NMP.
 72. The method of claim 71,wherein viscosity of the formulation is reduced by at least 10%.
 73. Themethod of claim 71, wherein viscosity of the formulation is reduced byat least 50%.
 74. The method of claim 71, wherein viscosity of theformulation is reduced by at least 80%.
 75. The method of claim 71,wherein the antibody is selected from the group consisting ofabagovomab, abciximab, actoxumab, adalimumab, afelimomab, afutuzumab,alacizumab, alacizumab pegol, ald518, alemtuzumab, alemtuzumab,alirocumab, altinumab, altumomab, amatuximab, anatumomab mafenatox,anrukinzumab, apolizumab, arcitumomab, aselizumab, atlizumab,atorolimiumab, bapineuzumab, basiliximab, bavituximab, bectumomab,belimumab, benralizumab, bertilimumab, besilesomab, bevacizumab,bezlotoxumab, biciromab, bivatuzumab, bivatuzumab mertansine,blinatumomab, blosozumab, brentuximab vedotin, briakinumab, brodalumab,canakinumab, cantuzumab mertansine, cantuzumab mertansine, caplacizumab,capromab pendetide, carlumab, catumaxomab, cc49, cedelizumab,certolizumab pegol, cetuximab, citatuzumab bogatox, cixutumumab,clazakizumab, clenoliximab, clivatuzumab tetraxetan, conatumumab,cr6261, crenezumab, dacetuzumab, daclizumab, dalotuzumab, daratumumab,demcizumab, denosumab, detumomab, dorlimomab aritox, drozitumab,duligotumab, dupilumab, ecromeximab, eculizumab, edobacomab,edrecolomab, efalizumab, efungumab, elotuzumab, elsilimomab,enavatuzumab, enlimomab pegol, enokizumab, enokizumab, enoticumab,enoticumab, ensituximab, epitumomab cituxetan, epratuzumab, erenumab,erlizumab, ertumaxomab, etaracizumab, etrolizumab, evolocumab,exbivirumab, exbivirumab, fanolesomab, faralimomab, farletuzumab,fasinumab, fbta05, felvizumab, fezakinumab, ficlatuzumab, figitumumab,flanvotumab, fontolizumab, foralumab, foravirumab, fresolimumab,fulranumab, futuximab, galiximab, ganitumab, gantenerumab, gavilimomab,gemtuzumab ozogamicin, gevokizumab, girentuximab, glembatumumab vedotin,golimumab, gomiliximab, gs6624, ibalizumab, ibritumomab tiuxetan,icrucumab, igovomab, imciromab, i mgatuzumab, inclacumab, indatuximabravtansine, infliximab, inolimomab, inotuzumab ozogamicin, intetumumab,ipilimumab, iratumumab, itolizumab, ixekizumab, keliximab, labetuzumab,lebrikizumab, lemalesomab, lerdelimumab, lexatumumab, libivirumab,ligelizumab, lintuzumab, lirilumab, lorvotuzumab mertansine,lucatumumab, lumiliximab, mapatumumab, maslimomab, matuzumab,mavrilimumab, mepolizumab, metelimumab, milatuzumab, minretumomab,mitumomab, mogamulizumab, morolimumab, motavizumab, moxetumomabpasudotox, muromonab-cd3, nacolomab tafenatox, namilumab, naptumomabestafenatox, narnatumab, natalizumab, nebacumab, necitumumab,nerelimomab, nesvacumab, nimotuzumab, nivolumab, nofetumomab merpentan,ocaratuzumab, ocrelizumab, odulimomab, ofatumumab, olaratumab,olokizumab, omalizumab, onartuzumab, oportuzumab monatox, oregovomab,orticumab, otelixizumab, oxelumab, ozanezumab, ozoralizumab,pagibaximab, palivizumab, palivizumab, panitumumab, panobacumab,parsatuzumab, pascolizumab, pateclizumab, patritumab, pemtumomab,perakizumab, pertuzumab, pexelizumab, pidilizumab, pintumomab,placulumab, ponezumab, priliximab, pritumumab, PRO 140, quilizumab,racotumomab, radretumab, rafivirumab, ramucirumab, ranibizumab,raxibacumab, regavirumab, reslizumab, rilotumumab, rituximab,robatumumab, roledumab, romosozumab, rontalizumab, rovelizumab,ruplizumab, samalizumab, sarilumab, satumomab pendetide, secukinumab,sevirumab, sibrotuzumab, sifalimumab, siltuximab, simtuzumab,siplizumab, sirukumab, solanezumab, solitomab, sonepcizumab, sontuzumab,stamulumab, sulesomab, suvizumab, tabalumab, tacatuzumab tetraxetan,tadocizumab, talizumab, tanezumab, taplitumomab paptox, tefibazumab,tefibazumab, telimomab aritox, telimomab aritox, tenatumomab,tenatumomab, teneliximab, teplizumab, teprotumumab, TGN1412,ticilimumab, tigatuzumab, tildrakizumab, TNX-650, tocilizumab,tocilizumab, toralizumab, tositumomab, tralokinumab, trastuzumab,TRBS07, tregalizumab, tremelimumab, tremelimumab, tucotuzumabcelmoleukin, tuvirumab, ublituximab, urelumab, urtoxazumab, ustekinumab,vapaliximab, vatelizumab, vedolizumab, veltuzumab, vepalimomab,vesencumab, visilizumab, volociximab, vorsetuzumab mafodotin, votumumab,zalutumumab, zanolimumab, zatuximab, ziralimumab, zolimomab aritox andthose shown in Table A.
 76. The method of claim 70, wherein the antibodyor fragment thereof is an anti-PCSK9 antibody.
 77. The method of claim76, wherein the antibody is evolocumab.
 78. A method for reducing theviscosity of a liquid pharmaceutical formulation comprising an antibodyor fragment thereof at a concentration of at least 140 mg/mL, comprisingthe step of combining the antibody or fragment thereof with aviscosity-reducing concentration of N-methyl pyrrolidone and N-acetylarginine.
 79. The method of claim 78, wherein viscosity of theformulation is reduced by at least 10%.
 80. The method of claim 78,wherein viscosity of the formulation is reduced by at least 50%.
 81. Themethod of claim 78, wherein viscosity of the formulation is reduced byat least 80%.
 82. The method of claim 78, wherein the antibody isselected from the group consisting of abagovomab, abciximab, actoxumab,adalimumab, afelimomab, afutuzumab, alacizumab, alacizumab pegol,ald518, alemtuzumab, alemtuzumab, alirocumab, altinumab, altumomab,amatuximab, anatumomab mafenatox, anrukinzumab, apolizumab, arcitumomab,aselizumab, atlizumab, atorolimiumab, bapineuzumab, basiliximab,bavituximab, bectumomab, belimumab, benralizumab, bertilimumab,besilesomab, bevacizumab, bezlotoxumab, biciromab, bivatuzumab,bivatuzumab mertansine, blinatumomab, blosozumab, brentuximab vedotin,briakinumab, brodalumab, canakinumab, cantuzumab mertansine, cantuzumabmertansine, caplacizumab, capromab pendetide, carlumab, catumaxomab,cc49, cedelizumab, certolizumab pegol, cetuximab, citatuzumab bogatox,cixutumumab, clazakizumab, clenoliximab, clivatuzumab tetraxetan,conatumumab, cr6261, crenezumab, dacetuzumab, daclizumab, dalotuzumab,daratumumab, demcizumab, denosumab, detumomab, dorlimomab aritox,drozitumab, duligotumab, dupilumab, ecromeximab, eculizumab, edobacomab,edrecolomab, efalizumab, efungumab, elotuzumab, elsilimomab,enavatuzumab, enlimomab pegol, enokizumab, enokizumab, enoticumab,enoticumab, ensituximab, epitumomab cituxetan, epratuzumab, erenumab,erlizumab, ertumaxomab, etaracizumab, etrolizumab, evolocumab,exbivirumab, exbivirumab, fanolesomab, faralimomab, farletuzumab,fasinumab, fbta05, felvizumab, fezakinumab, ficlatuzumab, figitumumab,flanvotumab, fontolizumab, foralumab, foravirumab, fresolimumab,fulranumab, futuximab, galiximab, ganitumab, gantenerumab, gavilimomab,gemtuzumab ozogamicin, gevokizumab, girentuximab, glembatumumab vedotin,golimumab, gomiliximab, gs6624, ibalizumab, ibritumomab tiuxetan,icrucumab, igovomab, imciromab, i mgatuzumab, inclacumab, indatuximabravtansine, infliximab, inolimomab, inotuzumab ozogamicin, intetumumab,ipilimumab, iratumumab, itolizumab, ixekizumab, keliximab, labetuzumab,lebrikizumab, lemalesomab, lerdelimumab, lexatumumab, libivirumab,ligelizumab, lintuzumab, lirilumab, lorvotuzumab mertansine,lucatumumab, lumiliximab, mapatumumab, maslimomab, matuzumab,mavrilimumab, mepolizumab, metelimumab, milatuzumab, minretumomab,mitumomab, mogamulizumab, morolimumab, motavizumab, moxetumomabpasudotox, muromonab-cd3, nacolomab tafenatox, namilumab, naptumomabestafenatox, narnatumab, natalizumab, nebacumab, necitumumab,nerelimomab, nesvacumab, nimotuzumab, nivolumab, nofetumomab merpentan,ocaratuzumab, ocrelizumab, odulimomab, ofatumumab, olaratumab,olokizumab, omalizumab, onartuzumab, oportuzumab monatox, oregovomab,orticumab, otelixizumab, oxelumab, ozanezumab, ozoralizumab,pagibaximab, palivizumab, palivizumab, panitumumab, panobacumab,parsatuzumab, pascolizumab, pateclizumab, patritumab, pemtumomab,perakizumab, pertuzumab, pexelizumab, pidilizumab, pintumomab,placulumab, ponezumab, priliximab, pritumumab, PRO 140, quilizumab,racotumomab, radretumab, rafivirumab, ramucirumab, ranibizumab,raxibacumab, regavirumab, reslizumab, rilotumumab, rituximab,robatumumab, roledumab, romosozumab, rontalizumab, rovelizumab,ruplizumab, samalizumab, sarilumab, satumomab pendetide, secukinumab,sevirumab, sibrotuzumab, sifalimumab, siltuximab, simtuzumab,siplizumab, sirukumab, solanezumab, solitomab, sonepcizumab, sontuzumab,stamulumab, sulesomab, suvizumab, tabalumab, tacatuzumab tetraxetan,tadocizumab, talizumab, tanezumab, taplitumomab paptox, tefibazumab,tefibazumab, telimomab aritox, telimomab aritox, tenatumomab,tenatumomab, teneliximab, teplizumab, teprotumumab, TGN1412,ticilimumab, tigatuzumab, tildrakizumab, TNX-650, tocilizumab,tocilizumab, toralizumab, tositumomab, tralokinumab, trastuzumab,TRBS07, tregalizumab, tremelimumab, tremelimumab, tucotuzumabcelmoleukin, tuvirumab, ublituximab, urelumab, urtoxazumab, ustekinumab,vapaliximab, vatelizumab, vedolizumab, veltuzumab, vepalimomab,vesencumab, visilizumab, volociximab, vorsetuzumab mafodotin, votumumab,zalutumumab, zanolimumab, zatuximab, ziralimumab, zolimomab aritox andthose shown in Table A.
 83. The method of claim 78, wherein the antibodyor fragment thereof is an anti-PCSK9 antibody.
 84. The method of claim83, wherein the antibody is evolocumab.
 85. A method for stabilizing aliquid pharmaceutical formulation comprising an antibody or fragmentthereof at a concentration of at least 140 mg/mL and N-methylpyrrolidone, comprising the step of combining the reduced viscosityliquid pharmaceutical formulation with a stabilizer selected from thegroup consisting of polysorbate 20, polysorbate 80, PLURONIC® F-68,lecithin, and protamine sulfate.
 86. A method for stabilizing a liquidpharmaceutical formulation comprising an antibody or fragment thereof ata concentration of at least 140 mg/mL and a combination of excipientscomprising N-methyl pyrrolidone and N-acetyl arginine, comprising thestep of combining the reduced viscosity liquid pharmaceuticalformulation with a stabilizer selected from the group consisting ofpolysorbate 20, polysorbate 80, PLURONIC® F-68, lecithin, and protaminesulfate.